Effects of nigral application of muscimol on release of [3H]γ-aminobutyrate and on multi-unit activity in various cat thalamic nuclei

The release of [³H]γ-aminobutyrate (GABA) neosynthesized from [³H]glutamine was estimated in one substantia nigra and in the ipsilateral thalamus of halothane-anesthetized cats by perfusing a [³H]glutamine-enriched physiological medium through a push-pull cannula implanted in the two structures under investigation. After two hours of superfusion, muscimol (10^?6 M) was delivered through the nigral push-pull cannula for 50–60 min and local- and distal-evoked changes of [³H]GABA release were analyzed. In some experiments, changes of global neuronal activity induced by muscimol application were recorded in different thalamic nuclei, using a bipolar electrode. In a few of the above experiments, biochemical and electrophysiological determinations were simultaneously performed in the substantia nigra and the thalamus. The nigral application of muscimol (10^?6 M) induced locally an activation of the substantia nigra reticulata cells, as well as an increase in release of [³H]GABA.Distally, in the thalamus, two types of biochemical and electrophysiological responses were observed according to the localization of the tip of the push-pull cannula or the electrode. (1) An increased release of [³H]GABA and a depression of the global multi-unit cellular activity were obtained in the ventralis medialis-ventralis lateralis, the centralis lateralis and the paracentralis nuclei. These effects could reflect an activation of the GABAergic nigrothalamic neurons projecting to these different thalamic nuclei. (2) In contrast, in the medialis dorsalis paralamellar zone adjacent to the intralaminar nuclei of the thalamus, a decrease of [³H]GABA release and an activation of the multi-unit activity were obtained. These latter results may suggest either a polysynaptic response or the non-GABAergic nature of the nigrothalamic neurons afferent to the medialis dorsalis paralamellar zone.     

Photodynamic sterilization of red cells and its effect on contaminating white cells: viability and mechanism of cell death

BACKGROUND: Phthalocyanines are useful sensitizers for photodynamic sterilization of red cell concentrates. Various lipid-enveloped viruses can be inactivated with only limited red cell damage. Because white cells are involved in the immunomodulatory effects of blood transfusions, the study of the effect of photodynamic treatment on these cells is imperative. STUDY DESIGN AND METHODS: White cell-enriched red cell suspensions were photodynamically treated with either the hydrophobic Pc4 (HOSiPcOSi-(CH3)2(CH2)3N(CH3)2) or water-soluble aluminum phthalocyanine tetrasulfonate (AIPCS4) under virucidal conditions. Viability of white cell subpopulations on Days 0, 1, and 4 after treatment was determined by fluorescence-activated cell sorting by flow cytometric analysis of propidium iodide uptake. Apoptosis induction was studied by DNA ladder formation and staining for an early marker of apoptosis (annexin V). RESULTS: Treatment with Pc4 causes a significant decrease in cell viability of all white cells, as shown by prodidium iodide uptake. Monocytes and granulocytes are the most sensitive, and lymphocytes are relatively more resistant. Some of the cells die by apoptosis, which is induced within 30 minutes after treatment. Treatment with AIPCS4 damages only monocytes; other cell populations are not affected. CONCLUSIONS: Physicochemical properties of the photosensitizers partly determine their effect on white cells. Differences in intracellular localization are likely to be responsible for the effects observed.     

夏氏疟原虫感染的红细胞膜表面抗原Pc90的cDNA特征 I．cDNA文库的构建及阳性克隆的筛选

目的：构建夏氏疟原虫早期滋养体的ｃＤＮＡ文库，并从中筛选出表达Ｐｃ９０的阳性克隆。方法：采用ＧＴＣ／ＣｓＣｌ法提取ＲＮＡ，ｃＤＮＡ克隆入λ－ＺＡＰ噬菌体，用抗Ｐｃ－ＨＣＰＭ血清３次筛选并经抗Ｐｃ９０单克隆抗体复筛，阳性克隆经体内剪接后行限制性内切酶酶切分析。结果：该ｃＤＮＡ文库有１．９×１０６个重组体。用抗Ｐｃ９０－ＨＣＰＭ的多克隆抗体筛选后，得到１６个阳性克隆，其长度分别为：０．６ｋｂｐ，１．４ｋｂｐ，１．６ｋｂｐ，２．２ｋｂｐ以及２．５ｋｂｐ。其中仅２．２ｋｂｐ和２．５ｋｂｐ克隆与抗Ｐｃ９０单克隆抗体反应呈阳性。限制性内切酶酶切分析表明，２．２ｋｂｐ，２．５ｋｂｐ和１．６ｋｂｐ的克隆具有不同的特征。结论：２．２ｋｂｐ和２．５ｋｂｐ的克隆极有可能表达Ｐｃ９０蛋白。     

Single thalamic neurons which project to both the rostral cortex and caudate nucleus studied with the fluorescent double labeling method

After injections of fast blue into the rostral cortex and Evans blue into the caudate nucleus in cats, doubly labeled neurons were present in the ventral anterior, ventral lateral, rhomboid, and mediodorsal thalamic nuclei. Doubly labeled cells were also found in most members of the intralaminar group, including the central medial, paracentral, central lateral, and parafascicular nuclei. Although the centromedian nucleus contained large numbers of cells labeled with Evans blue which project to the caudate nucleus, and a few fast-blue labeled cells which projected to the cortex, doubly labeled neurons were absent from this posterior intralaminar nucleus in this study.     

Projections of the zona incerta in the cat, with stimulation controls.

In eight cats single electrolytic lesions were placed in the zona incerta, and resultant fiber degeneration studies were made. In seven additional cats, stimulating electrodes were chronically implanted bilaterally into the zona incerta, H₂ (lenticular fasciculus), or H (prerubral) fields of Forel. The animals were placed in a two-compartment shuttle box, and a routinely established procedure of subthalamic stimulation was instituted. When the sensory (nociceptive) or motor manifestations and reactions were established, small lesions were made through both poles of the electrodes. The brains were studied by silver techniques for degenerating axons and terminals. Findings in the latter group of animals with physiologic substrates, compared to those in the first group, indicated that the zona incerta contains at least two major physiologic-anatomic components with differential fiber projections. The first component is a medial zona incerta proper or caudalis, paleospinothalamic, nociceptive-conducting system which causes typical escape responses. Its unequivocal projections are to the nucleus of the H₁ field, posterior and dorsal hypothalamus, part of the intralaminar system, ventromedial and ventralis anterior nuclei, nucleus reuniens, reticular nucleus, pulvinar, posterior nucleus, central gray, red nucleus, and the central tegmental tract. The second constitutent concerns pyramidal-extrapyramidal motor type responses that arise with avoidance reactions from other portions of the zona incerta. In these cases there is heavy projection to the caudate, entopenduncular, globus pallidus, and putamen nuclei. In contrast, degeneration from the nociceptive part of the zona incerta or H₂ and H fields to these nuclei is minimal.     

浙贝母素乙治疗大鼠卡氏肺孢子虫肺炎的实验研究

目的探讨浙贝母素乙注射液对大鼠卡氏肺孢子虫肺炎（PCP）的治疗作用,寻找杀灭或抑制卡氏肺孢子虫繁殖的药物,以提高包括艾滋病病人在内的免疫功能低下患者的生存质量。方法选择健康SD大鼠用地塞米松皮下注射,连续8周,每周2次,建立大鼠PCP模型;腹腔内注射不同剂量的浙贝母素乙注射液,连续5d,同时设有感染对照组和正常对照组。停药2周后杀鼠取肺,观察浙贝母素乙对大鼠肺内卡氏肺孢子虫发育的影响,肺组织印片检查包囊,肺组织病理切片染色观察组织变化。结果注射浙贝母素乙液注射组大鼠的肺组织印片包囊数量较治疗前明显减少,肺组织炎症反应明显好转。结论中药浙贝母素乙注射液对肺孢子虫在肺内发育有明显的抑制作用,有望用于PCP的治疗。     

Action monitoring in children with or without a family history of ADHD - Effects of gender on an endophenotype parameter

Attention-deficit/hyperactivity disorder (ADHD) is a frequent and highly heritable disorder overrepresented in boys. In a recent study investigating boys only, we found that action monitoring deficits as reflected by certain behavioral and electrophysiological parameters were familially driven. As gender may also have an important impact, this was examined in the current study with nonaffected children aged 8-15 years having relatives suffering from ADHD (N = 37, 21 ♀) and with age-matched controls without family history of ADHD (N = 33, 11 ♀).Extending our previous findings that action monitoring is a potential endophenotype for boys with ADHD, familially driven deficits were confirmed independently of gender. Thus, despite sharing the phenotype with controls, nonaffected siblings showed ADHD-like impairments albeit of smaller magnitude. However, girls performed generally more accurately, which in turn may have produced the differences between nonaffected siblings and controls in affective error processing that were not present in our boys-only assessment.     

Differences in regulatory T-cell and dendritic cell pattern in decidual tissue of placenta accreta/increta cases

Introduction

Primary infertility, miscarriage, and preeclampsia have been correlated with reduced numbers of regulatory T-cells (T_reg) suggesting that decreased extravillous trophoblast (EVT) invasion originates from inadequate EVT tolerance. In contrast increased numbers of T_reg-cells may be responsible for over-invasion of EVT. As the maturation status of dendritic cells (DC) influences T-cell behavior (tolerance or immune activation), altered relation between immature and mature DCs may also influence EVT invasion.

Method

Paraffin-embedded specimens of placenta accreta/increta (Pc; n = 11) and healthy intrauterine pregnancy (IUG; n = 18) were double-stained for cytokeratin and CD45, CD68, CD56, CD20, CD3, or CD8 as well as FoxP3/CD4 and FoxP3/CD8 and single-stained for CD4, CD25, FoxP3, CD209, Dec205 and CD83. Quantification of the leukocyte subpopulations was performed for decidua parietalis and basalis as characterized by cytokeratin-positive EVT. Statistical analysis was performed by using the Mann–Whitney test.

Result

There were significantly fewer CD4⁺ cells in Pc than in IUG. Concerning the T_reg-markers, FoxP3⁺ cells are significantly increased. CD25⁺ cells showed a small non-significant increase in Pc in comparison to IUG. Concerning dendritic cells, immature non-activated CD209⁺ DCs were significantly decreased in Pc while immature activated CD205⁺ DCs were slightly but non-significantly increased. Mature activated CD83⁺ DC were non-significantly decreased in IUG vs Pc.

Discussion and conclusion

The increased number of T_reg-cells in Pc suggests significance for these cells in the regulation of trophoblast invasion. Their adequate interaction with other lymphocyte populations (e.g. adequately maturated dendritic cells) may be one mechanism to assure controlled EVT invasion.