Exposure to cell phone radiation up-regulates apoptosis genes in primary cultures of neurons and astrocytes

The health effects of cell phone radiation exposure are a growing public concern. This study investigated whether expression of genes related to cell death pathways are dysregulated in primary cultured neurons and astrocytes by exposure to a working Global System for Mobile Communication (GSM) cell phone rated at a frequency of 1900MHz. Primary cultures were exposed to cell phone emissions for 2h. We used array analysis and real-time RT-PCR to show up-regulation of caspase-2, caspase-6 and Asc (apoptosis associated speck-like protein containing a card) gene expression in neurons and astrocytes. Up-regulation occurred in both "on" and "stand-by" modes in neurons, but only in "on" mode in astrocytes. Additionally, astrocytes showed up-regulation of the Bax gene. The effects are specific since up-regulation was not seen for other genes associated with apoptosis, such as caspase-9 in either neurons or astrocytes, or Bax in neurons. The results show that even relatively short-term exposure to cell phone radiofrequency emissions can up-regulate elements of apoptotic pathways in cells derived from the brain, and that neurons appear to be more sensitive to this effect than astrocytes.     

2型糖尿病相关基因的生物信息学分析

目的 通过生物信息分析途径,从分子水平揭示2型糖尿病的发病机制,为2型糖尿病的研究提供新的思路.方法 从公共数据库GEO中下载2型糖尿病相关基因芯片数据,利用Qlucore Omics Explorer 3.0软件筛选差异表达基因,STRING、DAVID等在线分析工具对差异表达基因进行下一步的生物信息学分析.结果 共筛选出89个差异基因,其中表达上调67个,下调22个,这些差异表达基因主要涉及到氧化还原反应、葡萄糖代谢过程、磷酸化作用、细胞骨架蛋白结合、核苷酸结合等分子功能和生物学过程.通过STRING分析,发现9个基因处在核心节点位置.结论 通过生物信息学的方法分析得出CDK9、TXN,NDUFS8基因可能为潜在的治疗靶点,需要下一步的分子生物学实验证实.     

Brain distribution of genes related to changes in locomotor activity

The relationship between genes and behavior, and particularly the hyperactive behavior, is clearly not linear nor monotonic. To address this problem, a database of the locomotor behavior obtained from thousands of mutant mice has been previously retrieved from the literature. Data showed that the percent of genes in the genome related to locomotor hyperactivity is probably more than 1.56%. These genes do not belong to a single neurotransmitter system or biochemical pathway. Indeed, they are probably required for the correct development of a specific neuronal network necessary to decrease locomotor activity. The present paper analyzes the brain expression pattern of the genes whose deletion is accompanied by changes in locomotor behavior. Using literature data concerning knockout mice, 46 genes whose deletion was accompanied by increased locomotor behavior, 24 genes related to decreased locomotor behavior and 23 genes not involved in locomotor behavior (but important for other brain functions) have been identified.These three groups of genes belonged to overlapping neurotransmitter systems or cellular functions. Therefore, we postulated that a better predictor of the locomotor behavior resulting from gene deletion might be the brain expression pattern. To this aim we correlated the brain expression of the genes and the locomotor activity resulting from the deletion of the same genes, using two databases (Allen Brain Atlas and SymAtlas). The results showed that the deletion of genes with higher expression level in the brain had higher probability to be accompanied by increased behavioral activity. Moreover the genes that were accompanied by locomotor hyperactivity when deleted, were more expressed in the cerebral cortex, amygdala and hippocampus compared to the genes unrelated to locomotor activity. Therefore, the prediction of the behavioral effect of a gene should take into consideration its brain distribution. Moreover, data confirmed that genes highly expressed in the brain are more likely to induce hyperactivity when deleted. Finally, it is suggested that gene mutations linked to specific behavioral abnormalities (e.g. inattention) might probably be associated to hyperactivity if the same gene has elevated brain expression.     

Antiviral NK cell responses in HIV infection: I. NK cell receptor genes as determinants of HIV resistance and progression to AIDS

NK cells play an important role in controlling viral infections. They can kill virus-infected cells directly as well as indirectly via antibody-dependent, cell-mediated cytotoxicity. They need no prior sensitization and expansion for this killing. NK cells are also considered as important regulators of antiviral immune responses. They do so by secreting a multitude of soluble mediators and by directly interacting with other immune cells, e.g., dendritic cells. NK cells do not possess a single well-defined receptor to recognize antigens on target cells. Instead, they express an array of inhibitory and activating receptors and coreceptors, which bind to their cognate ligands expressed on the surface of target cells. These ligands include classical and nonclassical MHC class I antigens, MHC-like proteins, and a variety of other self- and virus-derived molecules. They may be expressed constitutively and/or de novo on the surface of virus-infected cells. NK cell receptors (NKRs) of the killer-cell Ig-like receptor (KIR) family, like their MHC class I ligands, are highly polymorphic. Several recent studies suggest that epistatic interactions between certain KIR and MHC class I genes may determine innate resistance of the host to viral infections, including HIV. In the first part of this review article, we provide an overview of the current state of knowledge of NK cell immunobiology and describe how NKR genes, alone and in combination with HLA genes, may determine genetic resistance/susceptibilty to HIV infection and the development of AIDS in humans.     

Modulation of NK cell activity by CpG oligodeoxynucleotides

Oligodeoxynucleotides (ODN) with hypomethylated CpG motifs have been found to be potent stimulators of various aspects of innate and adaptive immunity. One of their major effects is the activation of natural killer (NK) killing activity in vitro and in vivo. There are several categories of CpG classified as type A, type B, and type C, although another category with inhibitory activity is being characterized further. CpG type A (CpG-A) is the most potent at activating NK cells. Examination of the cells and soluble mediators involved in this activation has led to an understanding of an interesting cascade of events. It appears that CpG activates dendritic cells (DC) which in turn activate NK-cells. This is not surprising since NK-cells do not seem to express TLR9, the CpG receptor. Of the various cytokines involved in NK-cell activation, it appears that type 1 interferon plays a pivotal role. Having activated NK-cells, DC themselves appear to become susceptible to lysis by the NK-cells they activated but with a delayed time kinetic. CpG ODN have been examined as monotherapeutic agents in murine tumor models. In one model, B16 melanoma, CpG ODN were very effective and NK cells were both necessary and sufficient for that effect. In another model, EL4 lymphoma, NK cells were necessary but not sufficient. Moreover, CpG were able to induce long-term survival in mice with established tumor.Studies in humans show similar results with potent activation in vitro. In a limited Phase I dose escalation study it also appeared that CpG ODN induce NK cell activation in humans in vivo.     

Defective NK cell activity following thermal injury.

Peripheral blood mononuclear lymphocytes (PBL) from thermal injury patients were examined for their ability to mediate natural killer (NK) cell activity against K562 tumour cells and against herpes simplex virus type 1 (HSV-1) infected Raji tumour cells. Using fluorescein isothiocyanate-conjugated monoclonal antibodies, the number of T3, T4, T8, Leu11, and Leu7 positive cells in PBL obtained from patients and normal controls was determined. Thermal injury patients had decreased levels of T3+ cells and a T4:T8 ratio which was significantly lower than that found in normal control individuals. Although patients had normal percentages of Leu7+ and Leu11+ cells, they had depressed NK cell activity against both K562 tumour cells and HSV-1 infected Raji cells. NK cell activity against K562 tumour cells was severely depressed during the first 20 days after injury. This defective NK cell activity did not appear to be due to a defect in PBL binding to the K562 tumour cells. In patients, the level of NK cell activity against HSV-1 infected cells did not correlate with the level of NK cell activity against K562 tumour cells. This finding further supports previous reports showing that NK cells which kill K562 tumour cells are different from the NK cell population which kills HSV-1 infected cells. Pretreatment of PBL obtained from patients with IL-2 or IFN-alpha, in some cases greatly enhanced NK cell killing of K562 tumour cells. However, IL-2 or IFN-alpha did not enhance NK cell activity in patients who had severely depressed levels of NK cell activity. Interestingly, in some patients, differential responsiveness to IL-2 and IFN-alpha was observed. In some patients, NK cell activity was enhanced by IL-2 but not by IFN-alpha. These results, while only suggestive, may indicate that different populations of NK cells respond preferentially to IL-2 and that IFN-alpha and/or IL-2 enhance NK cell activity in PBL obtained from some, but not all, thermal injury patients. Finally, this study clearly shows that thermal injury patients have defective NK cell activity not only against K562 tumour cells but also against virus-infected cells.     

Laughter regulates gene expression in patients with type 2 diabetes

BACKGROUND: Positive emotions influence endocrinological and immunological response. This study examined the effect of laughter,as an expression of positive emotion, in terms of gene expression changes. METHODS: Using a microarray technique, we analyzed the changes in expression of 18,716 genes from peripheral blood leukocytes in patients with type 2 diabetes, which were induced by laughter. RESULTS: Of the 18,716 genes, 23 genes showed significantly different expression changes after listening to the comic story compared to the lecture. Eight were relatively upregulated and 15 were downregulated 1.5 h after the laughing episode. However, these genes did not include genes that are directly involved in blood glucose metabolism. Among the 23 genes discriminated, all 4 genes encoding proteins involved in the immune response and all 4 signal transduction genes were downregulated. Moreover, it is noteworthy that 5 of the 8 relatively upregulated genes were related to the cell cycle, apoptosis, and cell adhesion. CONCLUSIONS: We demonstrated that laughter, which is an expression of positive emotion, is linked to gene expression. However, the finding of this study does not allow reasonable interpretation for the regulation of gene expression by laughter. A more focused study is needed that may identify the candidate genes for the association between physical condition and positive emotion.     

Interaction between immunoglobulin allotypes and NK receptor genes in diabetes post-hepatitis C virus infection

Genetic interactions between natural killer (NK) cells immunoglobulin-like receptor (KIR) genes and immunoglobulin allotypes have been previously reported in type 2 diabetes mellitus (DM) patients. Puerto Rican Americans with a history of intravenous drug use who developed DM following HCV infection (n = 32) were compared to individuals infected with HCV without diabetes (n = 121) and to DM non-infected individuals (n = 95). Subjects were genotyped for KIRs and immunoglobulin allotypes. We found interactions of immunoglobulin allotypes KM3/KM3 with NK inhibitory receptors 2DL3/2DL3, 2DL1 in the absence of 2DS4 associated with susceptibility to DM in HCV infected individuals. These data suggest the possibility that a subset of patients with HCV could have an immune-mediated component contributing to the development of DM.     

The evaluation of serial measurements of the NK cell activity in man

Studies on the NK cell activity require extensive controls because it is known that a series of variables can influence both the quality of effector cells and of target cells. This known variation of the susceptibility to lysis in one target cell line or even in sublines requires "standards", if separately performed experiments should be compared. In some laboratories, the experiments have been controlled for the variation of the target cell's susceptibility to lysis; others control for the differences of NK cell activities among subjects or for the variation of the NK cell activity in individuals from day to day. Each of these methods concentrates the main point of control to a particular variable. To demonstrate the power of several variables in relation to an induced and expected variation of the NK cell activity, we describe in this paper the results of a factorial analysis of variance testing PB-MNC from healthy donors against two target cells. The in vitro treatment of PB-MNC by IFN was chosen as induced variation of NK cell activity. As variables of the system, we defined subjects, days, target cells, and the effect of IFN. It is shown that the induced variable (IFN-treatment) counts for the most powerful variance component (32% of the total variance). Other large variance components are the differences of NK cell activities among subjects (20%), the day to day variation of the subjects' NK cell activities (13%), and the day to day variation of the susceptibility of target cells to lysis (13%). Since it is almost impossible to control for all variables in an experiment, some variables have to be selected which should be controlled for. The quantification of the variables, which is possible by a factorial analysis of variance, is useful for that purpose. This method of data analysis allows to estimate further the relevance of a particular expected effect if several variables, as in NK cell tests, influence the result of the experiment.     

单核细胞影响NK细胞抗K562细胞活性的实验研究

目的探讨单核细胞(MO)呼吸爆发过程中产生的反应性氮代谢物(RNM)、反应性氧代谢物(ROM)的情况及其对NK细胞抗K562细胞活性的影响。方法在NK+K562混合细胞培养体系中,观察加入MO或/和IL-2/PHA前后RNM、ROM产量、KIR及TNF-β、IFN-γ产量的相应变化。再观察加入二氢氯组胺、硫普罗宁后上述各指标的变化情况。结果①在NK+K562混合培养体系中,加入IL-2/PHA后,KIR逐渐升高;当按E/MO=10/2、10/5、10/10比例加入MO后,RNM、ROM产量随着MO数量的增加而增加,而TNF-β、IFN-γ的产量和KIR反而降低。对K562细胞数量作偏相关分析,RNM与KIR的负相关系数大于ROM。②加入药物后,ROM产量明显下降,KIR和TNF-β、IFN-γ产量明显升高,硫普罗宁还降低RNM的产量,但二氢氯组胺无效。结论①MO可通过产生ROM、RNM降低NK细胞的活性,抑制NK细胞抗K562细胞效应;②RNM对NK细胞的影响可能比ROM更强。③硫普罗宁通过清除ROM、RNM,可逆转MO对NK细胞的抑制作用。     

卵巢肿瘤患者外周血NK细胞受体NKG2A、NKG2D及NK活性检测的临床意义

通过研究卵巢癌及良性卵巢肿瘤患者外周血NK细胞表面受体的表达情况及NK活性的变化,分析探讨宿主NK细胞受体与肿瘤免疫逃逸的关系及其临床价值。分离受检者外周血单个核细胞,应用MTT法检测NK细胞的细胞毒活性,流式细胞术检测NK细胞受体NKG2D和NKG2A的表达,并结合临床病理因素作比较分析。结果显示,与良性卵巢肿瘤组和正常组相比,卵巢癌患者外周血NK细胞的细胞毒活性降低,NK细胞表面NKG2D的表达水平降低,而NKG2A的表达水平明显升高,其变化与卵巢癌的病情进展有关。此结果表明,卵巢癌患者机体NK细胞杀伤活性下降,NKG2D与NKG2A二者之间的平衡表达可能对NK细胞的功能状态起着重要的调节作用。     

控制自然杀伤细胞活性基因可能与HLA-B座位连锁

目的 研究中国人控制自然杀伤细胞（ｎａｔｕｒａｌ ｋｉｌｌｅｒ ｃｅｌｌ,ＮＫ）活性的基因是否与人类白细胞抗原等位基因相连锁。方法 ３４名健康无关个体,２５～６０岁,男１０名,女２４名,用血清学分型方法分析及其ＨＬＡ－Ａ－Ａ、Ｂ型。３１名家系成员,２２～７０岁,男１７名,女１４名,已经定出了ＨＬＡＩ、Ⅱ类及补体型,并已知其单倍型。用乳酸脱氢酶释放法测定上述对象的ＮＫ细胞活性。结果 聚类分析发现,     

Human NK cell response to pathogens

NK cells represent important effectors of the innate immunity in the protection of an individual from microbes. During an NK-mediated anti-microbial response, the final fate (survival or death) of a potential infected target cell depends primarily on the type and the number of receptor/ligand interactions occurring at the effector/target immune synapse. The identification of an array of receptors involved in NK cell triggering has been crucial for a better understanding of the NK cell biology. In this context, NCR play a predominant role in NK cell activation during the process of natural cytotoxicity. Regarding the NK-mediated pathogen recognition and NK cell activation, an emerging concept is represented by the involvement of TLRs and activating KIRs.NK cells express certain TLRs in common with other innate cell types. This would mean that specific TLR ligands are able to promote the simultaneous and synergistic stimulation of these innate cells, providing a coordinated mechanism for regulating the initiation and amplification of immune responses.Evidences have been accumulated indicating that viral infections may have a significant impact on NK cell maturation, promoting the expansion of phenotypically and functionally aberrant NK cell subpopulations. For example, during chronic HIV-infection, an abnormal expansion of a dysfunctional CD56neg NK cell subset has been detected that may explain, at least in part, the defective NK cell-mediated antiviral activity. An analogous imbalance of NK cell subsets has been detected in patients receiving HSCT to cure high risk leukemias and experiencing HCMV infection/reactivation. Remarkably, NK cells developing after CMV reactivation may contain “memory-like” or “long-lived” NK cells that could exert a potent anti-leukemia effect.     

体外抗原诱导T细胞对NK细胞功能的抑制作用及其影响因素

目的 研究①体外抗原诱导后,T细胞对NK细胞功能的抑制及其机制.②APC及不同T细胞亚群对这一过程的影响.方法 ①分离小鼠APC及T细胞,按1:3混合后分组进行诱导,48 h加入NK细胞通过51 Cr标准4 h释放实验观察NK细胞功能状况;激光共聚焦显微镜观察T细胞对NK细胞功能的抑制作用;RT-PCR法检测抗原诱导后小鼠T细胞bc1、bc2的表达.②标准51 Cr释放试验检测NK细胞功能以观察CD4 CD25 T细胞,抗原诱导的CD4 及CD4-T细胞对NK的抑制以及APC对体系中T-NK细胞间相互作用的影响.结果 ①51 Cr释放率分别为:Anti-CD80诱导组21%,抗原激活组24.4%,细胞对照组34%,无T细胞对照组32%,前3组上清液与NK细胞作用后靶细胞51 Cr释放率分别为34%、31.6%、33.1%.1、2两组51Cr释放率显著低于其余各组(P＜0.01),两组间无显著性差异.无T细胞对照组及各组上清液中51 Cr释放率无显著性差异.共聚焦显微镜下可观察到T细胞与NK细胞的直接接触.经抗原诱导48 h的细胞培养物可检出bc1、bc2的表达.②祛除或未祛除APC两组51 Cr的释放率均显著低于各对照组,且两组间无显著性差异(P＞0.05).CD4 CD25 T细胞组与CD4 T细胞组,51 Cr释放显著低于其他各组(P＜0.01),且前者显著低于后者(P＜0.05);CD4-T组与两对照组间无显著性差异.结论 ①抗原诱导T细胞对NK细胞的抑制作用可通过直接接触的方式,与共刺激因子CD80无关.T细胞上bc1、bc2表达是T细胞抑制NK细胞的可能分子机制之一;②抗原诱导的APC单独或通过T细胞均不能抑制NK细胞功能,提示T细胞对NK细胞的抑制是APC非依赖性的.抗原诱导后CD4 T细胞群体对NK细胞功能有显著抑制,而CD4-群体则无此特性.CD4 CD25 T细胞在无抗原参与情况下对NK细胞有显著性抑制作用.     

Laughter therapy modulates the parameters of renin-angiotensin system in patients with type 2 diabetes

The effect of laughter therapy on the plasma levels of renin, angiotensinogen, and prorenin was investigated in patients with type 2 diabetes. In the diabetic patients, the mean plasma renin concentrations were 24.6+/-12.1 ng/ml/h in the first observation (at the beginning of laughter therapy), 8.2+/-3.4 ng/ml/h in the second observation (three months after the beginning of laughter therapy) and 7.7+/-1.7 ng/ml/h in the third observation (six months after the beginning of laughter therapy). The mean plasma angiotensinogen concentrations in the 1st, 2nd and 3rd observations were 0.19+/-0.08, 0.47+/-0.12, 0.42+/-0.14 microg/ml, respectively. The mean plasma prorenin concentrations in the 1st, 2nd and 3rd observations during the laughter therapy were 195.1+/-66.2, 193.4+/-88.2 and 170.7+/-52.5 pg/ml, respectively. Plasma renin concentrations were significantly decreased (p<0.05) by the therapy. Subnormal concentrations of plasma angiotensinogen were found in the 1st observation and increased significantly (p<0.05) to the normal range after the therapy. Plasma prorenin concentration only slightly changed during the laughter therapy. Other biochemical parameters remained unchanged during the laughter therapy. These results indicated that a long-term laughter therapy changed the plasma components of renin-angiotensin system in patients with diabetes. Thus, laughter therapy can be used as non-pharmacological treatment for the prevention of diabetic microvascular complications.     

Cerebellar Purkinje cell activity related to the classically conditioned nictitating membrane response

Summary Because of the purported critical role of cerebellar lobule HVI in classical conditioning of the nictitating membrane response of the rabbit, we recorded extracellularly from HVI Purkinje cells (PCs) during differential conditioning. Rabbits were trained using tonal conditioned stimuli (CSs) and stimulation of the periocular region as the unconditioned stimulus (US). Many PCs responded to the US, the most frequently observed response being a burst of simple spikes. PCs in HVI showed a variety of responses to CSs that were related to conditioned responses (CRs). The most frequently observed response was an increase in simple spikes correlated with CRs. The activity of many of these cells antedated CRs by 20–200 ms. A smaller proportion of cells exhibited inhibition of simple spike activity that antedated CRs. The existence of PCs that alter their firing before CRs suggests that they may be causally involved in this behavior, and in this respect they reinforce reports that lesions of HVI or its connections disrupt nictitating membrane CRs. Although complex spike activity was not generally related to the US or to CRs, a few PCs responded in relation to CRs with only complex spikes. In demonstrating CR-related activity in cerebellar PCs, this study supports theories of cerebellar learning such as those of Marr and Albus.     

T cell responses to type 1 diabetes related peptides sharing homologous regions

Glutamic acid decarboxylase (GAD) 65 is a major autoantigen in type 1 diabetes. Regions of homology exist between GAD65 (residues 250-273) and the Coxsackie P2-C protein (residues 28-50) and between GAD65 (residues 506-518) and proinsulin (residues 24-36), and each of these has been reported to be a diabetes-associated T cell target. The aim of this study was to determine whether the homologous regions are shared targets of T lymphocyte reactivity in individual patients with type 1 diabetes. T cell proliferation against the corresponding peptide pairs, GAD254-276 and Coxsackie P2-C32-54 and GAD506-518 and proinsulin24-36, were measured in peripheral blood mononuclear cells from 26 patients with newly diagnosed type 1 diabetes and 24 control subjects. Responses with stimulation indices higher than 3 were found against each of the antigens tested in both patients and control subjects, and no differences were observed between groups. A strong positive correlation was found between responses to the corresponding peptide pairs GAD254-276 and Coxsackie P2-C32-54 (r=0.77, P<0.0001), and between responses to the corresponding peptide pairs GAD506-518 and proinsulin24-36 (r=0.66, P<0.0001). However, a similar correlation was also observed between responses to the noncorresponding pairs Coxsackie P2-C32-54 and proinsulin24-36 (r=0.82, P<0.0001), Coxsackie P2-C32-54 and GAD506-518 (r=0.82, P<0.0001), and GAD254-276 and proinsulin24-36 (r=0.83, P<0.0001). Strikingly, increased responses to peptides were found almost exclusively in subjects with high stimulation indices against the recall antigen tetanus toxoid, further suggesting that peripheral blood T cell responses are related to a general subject hyperreactivity. These data suggest that proliferative T cell responses to peptides containing putative autoreactive epitopes of GAD65 and proinsulin are not specific for type 1 diabetes, that correlation between T cell reactivity to peptides is not restricted to those containing homologous regions, and that non-antigen-specific factors are important determinants of in vitro measurements of T cell reactivity.