Recovery of in vitro functional activity of platelet concentrates stored at 4 degrees C and treated with second-messenger effectors

BACKGROUND: The potential for bacterial contamination limits the storage of platelets at 22 degrees C to 5 days. Refrigerated storage at 4 degrees C would abrogate this problem but would also result in a rapid loss of in vitro viability and functional activity and in vivo viability. The inhibition of platelets during storage by a combination of specific, reversible, second-messenger effectors has been investigated to allow prolonged storage at 4 degrees C with significant retention of in vitro viability and functional activity. STUDY DESIGN AND METHODS: The combination of effectors was added directly to platelet concentrates, and this step was followed by storage at 4 degrees C. Control units were incubated at 4 degrees C without the effectors and at 22 degrees C according to standard blood-banking techniques. At 1, 5, and 9 days, the units were tested for recovery of cell number, recovery of in vitro functional activity and viability, and expression of platelet surface markers. RESULTS: Treated platelets stored at 4 degrees C for 9 days, while spherical in shape, displayed no loss of cell number and had a recovery of viability and functional activity, as compared with control platelets stored at 22 degrees C for 5 days, as follows: ADP and collagen aggregation responses of 250 and 100 percent, respectively; a 70-percent recovery of hypotonic shock response; and a 60-percent recovery of extent of shape change. The treated platelets also expressed an equivalent amount of the surface marker glycoprotein lb and a lower amount of the activation marker alpha-granule membrane protein-140 on the membrane surface. CONCLUSION: Second-messenger effectors added to platelets significantly maintained in vitro functional activity with storage at 4 degrees C. In vitro analysis demonstrates the potential for extended 4 degrees C storage of platelets with numerical and functional recovery comparable to that achieved with current methods. Refrigerated storage of platelet concentrates has the potential to reduce the risk of bacterial contamination.     

Story stem responses of preschoolers with mood disturbances

Observing the young child's affect regulation and thought processes during a clinic assessment visit is of critical importance although challenging for children referred for mood disturbance. In this study, parents reported symptoms using standardized clinical interviews and story stems narratives were administered to 20 referred and 12 typically developing preschool age children. Comparison of the referred and typically developing children in our sample showed that specific story contexts varied in eliciting responses reflecting disorganization and thought disturbance from the referred children. The experience of using story stem narratives in the clinical assessment process suggests it provides a valuable complement to parent report for children referred for mood disturbance and mania symptoms but additional development and study of the method is necessary.     

蒙藏医传统药浴超微粉中槲皮素的定性与定量研究

目的:建立传统药浴超微粉中槲皮素的定性鉴别与含量测定方法,对其进行定性与定量研究。方法:采用薄层色谱法(TLC)对传统药浴超微粉中槲皮素进行定性鉴别,并采用高效液相色谱法(HPLC)对其进行含量测定。色谱柱为pgrandil STC-C18(4.6mm×250mm,5μm),以甲醇(A)-0.4%磷酸水(B)为流动相,梯度洗脱,检测波长为360nm,流速1.0m L/min,柱温30℃,进样量10μL。结果:经薄层色谱法鉴别,槲皮素对照品与样品处显同一斑点。槲皮素在0.09296μg~0.46480μg范围内,呈良好的线性关系(r=0.9999),平均回收率为96%,RSD=1.51%,n=10。结论:本法准确、简便,具有良好的重复性,可为传统药浴超微粉质量控制提供依据。     

艾灸对阳虚体质者任脉热学特性的影响*

目的 观察艾灸神阙和关元对阳虚体质者任脉及其旁开组织皮肤温度的影响,探讨艾灸对阳虚体质的作用机制。方法 将31名阳虚体质志愿者随机分为对照组（n=15）和艾灸组（n=16）,对照组进行健康宣教,艾灸组进行健康宣教和艾灸神阙、关元穴干预,分别于干预前后运用红外热成像仪记录任脉穴位、任脉(胸腹段)及其左右旁开1.5寸非经对照线的体表温度。结果 1. 与干预前比较,艾灸组受试者任脉及其左右旁开1.5寸非经对照线体表平均温度均升高,其中任脉及其左旁开呈明显升高（P<0.05,P<0.05）,任脉线上体表平均温度高于其左右旁开非经对照线（P<0.05,P<0.01）,左旁开对照线平均温度明显高于右旁开对照线（P<0.05）;对照组干预前后无明显差异。2. 与干预前比较,艾灸组受试者任脉线上天突穴（P<0.05）、膻中穴（P<0.05）体表温度均明显升高,中脘穴和气海穴体表温度升高,但尚未达统计学差异。对照组干预前后任脉线上穴位体表温度均无统计学差异。3. 干预前两组受试者任脉线上穴位温度均呈自上而下逐渐降低趋势,干预后艾灸组各穴位温度均上升,胸段穴位升高较为显著,总体上仍呈上热下寒趋势。结论 艾灸神阙和关元可以升高任脉经穴及其旁开组织体表温度,对机体起整体性调节作用,但短期内尚未改变其上热下寒的温度趋势;艾灸神阙和关元对任脉经穴及其旁开部位的升温程度差异可能与各部位的相对阴阳属性有一定关系。     

Cytokine production in IgG-mediated red cell incompatibility

The transfusion of incompatible red cells may result in fever and systemic symptoms. The mechanisms by which these symptoms are produced in the setting of antibodies that do not usually fix complement, as in the Rh system, are obscure. It has been hypothesized, on the basis of their known biologic activities, that a specific set of cytokines may be involved in such transfusion reactions. Therefore, the production of the inflammatory cytokines interleukin-1 beta (IL-1 beta), tumor necrosis factor (TNF), interleukin-6 (IL-6), and interleukin-8 (IL-8) by human monocytes in response to red cells sensitized with anti-D was investigated, as a model of IgG-dependent hemolytic transfusion reactions. IL-1 beta, IL-6, and IL-8 were detectable in the culture supernatants at 4 to 6 hours and increased up to 24 hours, whereas TNF peaked at 6 hours. Immunocytochemical stains of cell preparations demonstrated IL-1 beta, IL-8, and TNF in monocytes engaged in erythrophagocytosis. IL-8 production and phagocytosis could be inhibited by monomeric IgG, but Fab fragments of a monoclonal antibody specific for the low-affinity IgG receptor Fc gamma RII could not be, which suggests the involvement of the high-affinity receptor Fc gamma RI. Neutralizing antisera to IL-1 beta and TNF did not abrogate the production of IL-8, which suggests that sensitized red cells serve as a primary signal for this cytokine. These findings indicate that the production of inflammatory cytokines by phagocytes may be responsible for the symptomatology of IgG-mediated hemolytic transfusion reactions.     

GSK189254, a novel H3 receptor antagonist that binds to histamine H3 receptors in Alzheimer's disease brain and improves cognitive performance in preclinical models

6-[(3-Cyclobutyl-2,3,4,5-tetrahydro-1H-3-benzazepin-7-yl)oxy]-N-methyl-3-pyridinecarboxamide hydrochloride (GSK189254) is a novel histamine H(3) receptor antagonist with high affinity for human (pK(i) = 9.59 -9.90) and rat (pK(i) = 8.51-9.17) H(3) receptors. GSK189254 is >10,000-fold selective for human H(3) receptors versus other targets tested, and it exhibited potent functional antagonism (pA(2) = 9.06 versus agonist-induced changes in cAMP) and inverse agonism [pIC(50) = 8.20 versus basal guanosine 5'-O-(3-[(35)S]thio)triphosphate binding] at the human recombinant H(3) receptor. In vitro autoradiography demonstrated specific [(3)H]GSK189254 binding in rat and human brain areas, including cortex and hippocampus. In addition, dense H(3) binding was detected in medial temporal cortex samples from severe cases of Alzheimer's disease, suggesting for the first time that H(3) receptors are preserved in late-stage disease. After oral administration, GSK189254 inhibited cortical ex vivo R-(-)-alpha-methyl[imidazole-2,5(n)-(3)H]histamine dihydrochloride ([(3)H]R-alpha-methylhistamine) binding (ED(50) = 0.17 mg/kg) and increased c-Fos immunoreactivity in prefrontal and somatosensory cortex (3 mg/kg). Microdialysis studies demonstrated that GSK189254 (0.3-3 mg/kg p.o.) increased the release of acetylcholine, noradrenaline, and dopamine in the anterior cingulate cortex and acetylcholine in the dorsal hippocampus. Functional antagonism of central H(3) receptors was demonstrated by blockade of R-alpha-methylhistamine-induced dipsogenia in rats (ID(50) = 0.03 mg/kg p.o.). GSK189254 significantly improved performance of rats in diverse cognition paradigms, including passive avoidance (1 and 3 mg/kg p.o.), water maze (1 and 3 mg/kg p.o.), object recognition (0.3 and 1 mg/kg p.o.), and attentional set shift (1 mg/kg p.o.). These data suggest that GSK189254 may have therapeutic potential for the symptomatic treatment of dementia in Alzheimer's disease and other cognitive disorders.     

Lymphoblastic lymphoma in adults: results of a pilot protocol

Thirteen adult patients with histologically confirmed lymphoblastic lymphoma were treated with an intensive chemotherapy program consisting of induction with cyclophosphamide, adriamycin, vincristine, and prednisone (modified CHOP); consolidation and central nervous system (CNS) prophylaxis with methotrexate intrathecally and by high-dose intravenous injection, citrovorum factor and L-asparaginase; reinforcement with CHOP; and maintenance with 6-mercaptopurine and methotrexate. Treatment duration was 1 yr. A 14th patient with T-cell acute lymphoblastic leukemia was also treated at presentation by the same regimen. Thirteen patients had at least a mediastinal mass or abnormal cells in the bone marrow; one presented with CNS disease. The median age was 22 yr (range 16--50), and male--female ratio was 2.5:1. All patients had a rapid complete clinical response. Of the 13 patients without initial CNS disease, 4 have relapsed, 3 with primary CNS relapse and 1 with a recurrent abdominal mass. Five patients have died, 2 from drug toxicity, 2 from CNS relapse, and 1 from chronic myelogenous leukemia, which was diagnosed simultaneously with the lymphoblastic lymphoma. The median follow-up is 19 mo, and all patients have completed their planned therapy. At 3 yr, the actuarial survival is 61% and relapse-free survival is 56%.     

Quadricep ACL Reconstruction Techniques and Outcomes: an Updated Scoping Review of the Quadricep Tendon

Purpose of ReviewThe purpose of this review is to provide an up-to-date summary on the most recent literature examining techniques and outcomes in anterior cruciate ligament (ACL) reconstruction using quadriceps tendon (QT) which will enable surgeons to make well informed evidence-based decisions when choosing a particular graft option and technique in ACL reconstruction.Recent FindingsSeveral RCTs and systematic reviews have been published recently on this topic, and overall, there were no differences found between the QT, HT, and BPTB groups in patient-reported outcomes, stability testing, or graft re-rupture rates. In terms of strength testing, the QT group did have inferior knee extensor strength on isokinetic testing when compared to the HT group, whereas the HT group had inferior knee flexor strength compared to the QT group. No differences were found on strength testing between the QT and BPTB groups. Currently, two large RCTs, the Stability2 and SQuASH trials, are ongoing examining the effectiveness of the QT vs BPTB with or without LET and QT vs HT in the pediatric population which will help shed further light on the effectiveness of the QT as a graft choice in ACL reconstruction.SummaryThe findings of this scoping review demonstrate that the QT is an excellent graft option in ACL reconstruction both in the primary and revision settings, among adult and pediatric populations. This review provides surgeons with further assurance when selecting QT autograft in ACL reconstruction.     

消化系恶性肿瘤患者血清与腹水中细胞因子活性变化

目的研究消化系恶性肿瘤（ＤＭＴ）患者血清与腹水中内源性ＩＬ２,ＩＬ６,ＩＬ８,ＴＮＦα和ＩＦＮγ的生物学活性．方法应用ＥＬＩＳＡ法检测了１５例ＤＭＴ患者（肝癌１１例,胆总管癌１例,胰腺癌１例,胃癌１例,直肠癌１例）血清与腹水中５种细胞因子活性,并与６例肝硬变（ＬＣ）患者和８例正常成人进行了比较分析．结果ＤＭＴ患者血清ＩＬ２,ＩＬ６的生物学活性显著低于ＬＣ（Ｐ＜００５）;腹水中ＩＬ２,ＩＬ８活性显著低于ＬＣ组（Ｐ＜００１）,而ＩＬ６和ＩＦＮγ活性则高于ＬＣ组（Ｐ＜００１,００５）．ＤＭＴ患者血清中ＩＬ６,ＩＬ８活性明显高于正常成人组（Ｐ＜００５）;ＩＬ２,ＩＦＮγ则低于正常成人组,但缺乏显著性．肝癌血清和腹水中ＩＬ２活性显著高于非肝癌组（Ｐ＜００５）;而ＩＬ６活性则相对降低（Ｐ＜００５）．结论恶性肿瘤患者血清中ＩＬ２和ＩＦＮγ活性低于正常人,是ＤＭＴ患者抗肿瘤免疫功能缺陷的标志．ＩＬ６对于预测ＤＭＴ患者的预后具有重要的意义     

Granzyme A loading induces rapid cytolysis and a novel form of DNA damage independently of caspase activation.

Cytotoxic lymphocytes trigger apoptosis by releasing perforin and granzymes (Grn). GrnB activates the caspase apoptotic pathway, but little is known about GrnA-induced cell death. Perforin was used to load recombinant GrnA and GrnB and enzymatically inactive variants into target cells. GrnA induces single-strand DNA breaks that can be labeled with Klenow polymerase and visualized on alkaline gels. GrnA-induced DNA damage but not cytolysis requires GrnA proteolysis. GrnA-induced membrane perturbation, nuclear condensation, and DNA damage are unimpaired by caspase blockade. GrnA fails to induce cleavage of caspase-3, lamin B, rho-GTPase, or PARP. GrnA-induced cytotoxicity and cleavage of PHAP II, a previously identified GrnA substrate, are unimpaired in Jurkat cells that overexpress bcl-2. Therefore, GrnA activates a novel apoptotic pathway.