Immunohistochemical and immunoelectron microscopical characterization of cerebrovascular and senile plaque amyloid in aged dogs'' brains

Immunohistochemical and immunoelectron microscopical studies were carried out on 28 aged dogs' brains. Amyloid deposits were seen in the arteries and capillaries in the leptomeninges and in superficial areas of the cortices in 19 (67.9%) of the 28 dogs (10-22 years of age). Immunohistochemically, these amyloid deposits were reactive for anti-beta/A4 antibody. Additionally, a variable number of parenchymal deposits with diffuse beta/A4-immunoreactivity (diffuse plaques) was also noted throughout the cerebral cortex in 24/28 dogs (85.7%). However, these plaque lesions were undetectable in Congo red staining. Electron microscopically, amyloid fibrils, measuring 10 nm in width, were located mainly in the tunica media of the arteries, and in less involved vessels they tended to be present among collagen fibres in the adventitia and smooth muscle cells in the outer layer of the media. The plaque lesions appeared to contain sparse aggregations of amyloid fibrils. In immunoelectron microscopical examinations, all amyloid fibrils in both blood vessels and plaques were selectively labelled by gold particles. These findings indicate that aged dogs can provide a useful experimental model for research into the beta/A4-type of cerebral amyloidosis commonly seen in Alzheimer's disease.     

pp60c-src encoded by the proto-oncogene c-src is a product of sensory neurons

The advent of recombinant DNA technology has led to the identification in the DNA of normal animal cells of over 30 proto-oncogenes that are homologous to retroviral transforming genes. One of these encodes a protein kinase (pp60c-src) of unknown function, that is preferentially synthesized in brain and neural retina. Here the expression of pp60c-src in the peripheral nervous system was examined in sensory neurons from chick dorsal root ganglia with antisera raised against the transforming protein of Rous sarcoma virus (pp60v-src) expressed in Escherichia coli carrying the cloned v-src gene. This antiserum recognizes pp60c-src specifically in normal chicken cells. Western immunoblotting showed that dorsal root ganglia of stage 30 (day 6.5) chick embryos contained elevated levels of pp60c-src. Immunoperoxidase staining of neuron-enriched cultures prepared from chick dorsal root ganglia showed pp60c-src immunoreactivity in cells with neuronal morphology; flat, fibroblastic cells contained no detectable immunoreactivity. Indirect double immunofluorescence with pp60src antibodies and monoclonal antibodies against the 200-kD subunit of neurofilament protein confirmed that the cells expressing pp60c-src were neurons. Ninety-six percent of the neurofilament-positive cells were immunoreactive with pp60src antibodies, and conversely, all pp60c-src-positive cells were immunoreactive with neurofilament antibodies. pp60c-src immunofluorescence appeared to be distributed over the cell body, processes, and growth cones. These results clearly demonstrate that pp60c-src is a product of neurons and is expressed in sensory neurons in culture.     

卡维地洛对不稳定型心绞痛患者血浆炎症因子的影响

目的观察卡维地洛对不稳定型心绞痛（UAP）患者高敏C反应蛋白（hs-CRP）、白介素-6（IL-6）及细胞间可溶性黏附分子1（sICAM-1）水平的影响。方法UAP患者62例采用完全随机化方法分成对照组（n=30）和治疗组（n=32）,在常规抗血小板、扩血管治疗基础上,对照组予美托洛尔（12.5mg,2次/d×3d）口服,治疗组服卡维地洛（6.25mg,2次/d×3d）,在治疗前后分别测定hs-CRP、IL-6、sICAM-1值。结果对照组和治疗组在治疗前hs-CRP、IL-6、sICAM-1均无显著性差异;用药后两组3指标均较用药前显著降低（P<0.05）;治疗组在用药后IL-6、sI-CAM-1显著低于对照组（P<0.05）。同时,用药后两组患者的心率、血压、心肌耗氧量均较用药前显著降低（P<0.05）,治疗组的心肌耗氧量显著低于对照组（P<0.05）。结论卡维地洛可显著降低UAP患者的炎症因子IL-6、sICMA-1水平。     

Effect of divalent cations on structure-function relationships of the antitumor protein α-sarcin

α-Sarcin binds one Zn(II) cation per protein molecule, with a K_d value of 0.9 mM, determined by equilibrium dialysis experiments. Ca(II), Mg(II), and Mn(II) do not bind to α-sarcin. Cd(II) and Co(II) also behave as Zn(II). The binding produces local modifications on the protein conformation affecting the microenvironment of tryptophan residues. The three cations modify the fluorescence emission of the protein. The near-u. v. circular dichroism spectrum of the protein is also altered. The binding of Zn(II) and related cations does not modify the secondary structure of the protein. The ribonucleolytic activity of a-sarcin is inhibited upon Zn(II) binding, but no alteration of the ability of the protein to aggregate phospholipid vesicles has been observed.     

Parvalbumin-immunoreactive neurons in the cerebral cortex of the lizardPodarcis hispanica

An antibody against the calcium binding protein parvalbumin selectively labels a set of neurons in the cerebral cortex of lizards. Golgi-like immunostained bipolar, multipolar and pyramid-like neurons appear mainly located in the inner plexiform layers. Parvalbumin-immunoreactive (PARV-IR) puncta are concentrated in the cell layer of the dorsal and dorsomedial cortices showing a basket-like distribution. The morphology and distribution of PARV-IR neurons and puncta overlap GABA-immunostaining in the cerebral cortex of lizards. Thus, it is likely that PARV-IR neurons are a subset of the cortical GABAergic neurons of lizards.     

瘦素、C-反应蛋白与老年急性冠脉综合征患者的相关研究

目的探讨急性冠脉综合征(ACS)患者血浆瘦素(LP)及C-反应蛋白(CRP)水平的变化及其临床意义。方法测定急性心肌梗死组(AMI组)、不稳定型心绞痛组(UA组)、对照组(健康体检者)血浆LP及CRP浓度,分析LP、CRP与病情严重程度的关系以及LP与CRP的相关性。结果AMI组、UA组患者血浆LP、CRP浓度较对照组显著升高(P<0.01),2组患者血浆LP与CRP浓度之间均呈显著正相关;AMI组血浆LP、CRP浓度比UA组明显升高(P<0.05)。结论血浆LP及CRP浓度与ACS病情严重性呈正相关,且LP与CRP浓度呈正相关。说明LP与CRP共同参与组织炎症反应,是ACS重要的危险因素及预测因子。     

高+Gz重复暴露对大鼠脑组织胶质纤维酸性蛋白表达的影响

目的观察高＋Gz重复暴露后脑组织星形胶质细胞中胶质纤维酸性蛋白（GFAP）表达的变化。方法SD大鼠30只，随机分为对照组和＋10G；暴露组。＋G2暴露组的大鼠暴露于＋10Gz／45S，共暴露5次，中间间隔5min。分别于＋Gz暴露后不同时间灌注取脑，免疫组织化学染色观察脑GFAP的表达情况。结果＋Gz暴露后1d、2d，顶叶皮层、海马GFAP阳性细胞数较对照组均显著增多（P〈0．05），以弱阳性细小型为主；暴露后4d、6d，顶叶皮层、海马GFAP阳性细胞数较对照组进一步增多（P〈0．01），以中等阳性细胞为主，肥大的GFAP增多。结论重复暴露＋10Gz／45s可引起大鼠顶叶皮层、海马GFAP表达显著增加。     

Bacterial ghosts (BGs)—Advanced antigen and drug delivery system

Bacterial ghosts (BGs) are empty bacterial envelopes of Gram-negative bacteria produced by controlled expression of cloned gene E, forming a lysis tunnel structure within the envelope of the living bacteria. BGs are devoid of cytoplasmic content and possess all bacterial bio-adhesive surface properties in their original state while not posing any infectious threat. BGs are ideally suited as an advanced drug delivery system (ADDS) for toxic substances in tumor therapy. The inner space of BGs can be loaded with either single components or combinations of peptides, drugs or DNA which provides an opportunity to design new types of (polyvalent) drug delivery vehicles. Uptake of BGs loaded with Doxorubicin (Dox) by CaCo2 cells led to effective Dox release from endo-lysosomal compartments and accumulation in the nucleus. Viability and proliferative capacity of the cells were significantly decreased (2–3 orders of magnitude) after internalization of Dox loaded BGs as compared to cells incubated with free Dox. The same effect was observed with leukemia cells. Melanoma cells also revealed a high capability to internalize BGs. These results indicate that BGs are able to target a range of types of cancer. BGs have also been investigated as DNA delivery vectors. Studies show DNA loaded BGs are efficiently phagocytosed and internalized by both professional APCs and tumor cells with up to 82% of cells expressing the plasmid-encoded reporter gene. Our studies with BGs as an ADDS system contribute (i) to optimize drug delivery for the treatment of cancer; (ii) define specific conditions for selection and preparation of BG formulations; (iii) and provide a background for the clinical application of BGs in cancer therapy.