Activation of antifreeze proteins from larvae of the beetle Dendroides canadensis

Summary Purified antifreeze proteins (AFPs) from the larvae of the beetle Dendroides canadensis do not produce the high levels of antifreeze activity seen in the hemolymph of overwintering larvae, even when the purified AFPs are assayed at very high concentrations. However, addition of certain proteins or agar (at concentrations sufficiently low that the gel state does not result) to the Dendroides AFP resulted in a 2–3-fold increase in activity. A 70-kDa protein with AFP-activating capabilities was purified from Dendroides larvae. Addition of this endogenous activator protein to a 4 mg·ml^-1 solution of AFP increased the activity of the AFPs to values comparable to those of the hemolymph of overwintering larvae. Data derived from a modified immunoblot technique demonstrate that the activators bind to the AFP, or vice versa. Formation of this association must allow the AFP to block ice crystal growth by binding to the surface of potential seed crystals in the normal fashion. However, because the AFP-activator complex is much larger than the AFP alone, the complex probably blocks a greater surface area of the crystal and is thus a more efficient antifreeze.Abbreviations AFP antifreeze protein - BSA bovine serum albumine - DEAE diethylaminoethyl - Ig immunoglubolin - LPIN lipoprotein ice nucleator - PIN protein ice nucleator - SDS sodium dodecyl sulfate - PAGE polyacrylamide gel electrophoresis - TH thermal hysteresis     

Immunopathological features of human pulmonary tumors following low-dose interleukin-2

Summary We administered preoperative low-dose interleukin-2 (IL-2) to 10 patients undergoing thoracotomy for pulmonary tumors. The in vivo effect of IL-2 on tumor-associated lymphocyte activity was assessed in the resected specimens by immunohistochemistry and compared with observations in 45 patients who did not receive IL-2. H & E evaluation revealed an increase in intra- and peritumoral lymphocyte infiltration in the IL-2-treated patients. Immunopathological evaluation with monoclonal antibodies revealed that this lymphocyte infiltration was predominantly CD5-positive T cells. The amount of intra-and peritumoral lymphocyte activity correlated with the dose of IL-2 administered (6000–90 000 international units/kg every 8 h for 48 h. IL-2-treated patients showed increases in T-cell-associated activation markers (IL-2 -receptor, transferrin receptor and HLA-DR) on peritumoral lymphocytes, but not on intratumoral lymphocytes. We previously reported that low-dose IL-2 increases the intrinsic natural killer cell cytotoxicity of intratumoral lymphocytes and suggest that this lymphocyte infiltration is further evidence that low-dose IL-2 can augment in vivo lymphocyte activity at the tumor site.This work was supported in part by USPHS grants CA 44 352 (S. H. G.) and 43 658 (A. J. C.). S. G. S. was supported by NIH Surgical Oncology Training Grant CA 09 010     

Characterization of a monoclonal antibody that binds equally to all apolipoprotein and lipoprotein forms of human plasma apolipoprotein B. II. Isolation of apolipoprotein B-containing lipoproteins from human plasma

Monoclonal antibody ('Pan B' antibody) that binds equally to all major forms of human plasma apolipoprotein B was used in an immunoaffinity chromatography procedure to isolate apolipoprotein B-containing lipoproteins from hyperlipidemic human plasma. These lipoproteins were compared with lipoproteins in native plasma, with lipoproteins isolated by polyclonal antibodies and with lipoproteins isolated by the conventional ultracentrifugational method. Judged by the apolipoprotein and lipid composition, lipoproteins isolated with 'Pan B' antibody were virtually identical to those isolated by ultracentrifugation or polyclonal antibodies. Lipoproteins isolated by 'Pan B' antibody were comparable in size and shape to the lipoproteins in native plasma and to the lipoproteins isolated by polyclonal antibodies or ultracentrifugation. The immunoaffinity column with monoclonal 'Pan B' antibody retained all apolipoprotein B-containing lipoproteins and showed significantly higher capacity than polyclonal immunoaffinity column. The column with the highest capacity allowed the isolation from whole plasma of 0.144 mg of apolipoprotein B per ml of gel in less than 2 h.     

Anisodamine induces the hexagonal phase in phospholipid liposomes

The effects of anisodamine on the polymorphic phase behaviour of cardiolipin and dioleoylphosphatidylcholine liposomes have been investigated by freeze-fracture electron microscopy. Anisodamine induces the formation of lipidic particles in cardiolipin liposomes at pH 7.0 and hexagonal HII tubes at pH 8.8. When the molar ratio of anisodamine and dioleoylphosphatidylcholine is 4 to 1, lipidic particles can be observed in the fracture faces.     

Histidyl-tRNA synthetase, the myositis Jo-1 antigen, is cytoplasmic and unassociated with the cytoskeletal framework

The myositis-specific anti-Jo-1 autoantibody, which is directed against histidyl-tRNA-synthetase, is found in 30% of polymyositis patients. The Jo-1 antigen has been reported to be a nuclear antigen by some authors. On the contrary we show that less than 2% of the total histidyl-tRNA and lysyl-tRNA synthetase activities are associated with purified rat liver nuclei or the hepatocyte intermediate filament-nuclear fraction. In the presence of polyethylene glycol, in which the high Mr multi-enzyme complex containing lysyl-tRNA synthetase is insoluble, 65% of the lysyl-tRNA synthetase and only 15% of histidyl-tRNA synthetase activities remained associated with the cytoskeletal framework. The Jo-1 antigen exhibited a diffuse granular cytoplasmic distribution in cultured rat hepatocytes as determined by indirect immunofluorescent microscopy. Hence, the Jo-1 antigen is cytoplasmic and unassociated with the cytoskeletal framework or high Mr synthetase complex in situ.     

Studies on the phospholipid composition of pathogenic cell membranes of Mycoplasma hyopneumoniae

The membrane phospholipid and fatty acid compositions of Mycoplasma hyopneumoniae, a pathogen of porcine enzootic pneumoniae isolated in China, was studied by thin-layer chromatography and gas chromatography. The results showed that membrane phospholipids consisted predominantly of diphosphatidylglycerol. The percentage of C16 - C18 fatty acids comprised 79% of the total fatty acids, of which oleic acid as well as palmitic acid are the major fatty acids. Some differences were shown in fatty acid composition as compared with membranes of other species of Mycoplasma.     

A comparative study of the tapetum, retina and skull of the ferret, dog and cat

Results of this investigation indicate that the ferret (Mustela putorius) closely resembles the dog (Canis familiaris) and should be a useful research animal alternative. The tapetum lucidum is a common structure present in the eyes of dogs, cats (Felis catus) and other nocturnal animals. Our study showed that this structure was present in ferret eyes. The color or reflection of the ferret and dog tapetum was remarkably reduced by the general fixation with glutaraldehyde. However, this color fading phenomenon was not observed in the cat tapetum. These observations led to this comparative study on several morphological, histochemical and biochemical parameters on mature ferrets, dogs and cats including: (1) the number of center tapetum cell layers, (2) thickness of center tapetum, (3) presence of a microtubule-like structure in each tapetal rod, (4) presence of electron-dense cores in tapetal rods after prolonged fixation in glutaraldehyde, (5) retention of reflection or color of tapetum after prolonged glutaraldehyde fixation, (6) zygomatic bones of eye orbits, (7) zinc content in tapetum, (8) cysteine in the tapetum, (9) cysteine sulfinic acid decarboxylase in liver, (10) thickness of retina from center tapetum, (11) anterior view of skull configuration, and (12) lateral view of skull configuration (jaw and teeth). Among these 12 parameters, ferret and dog were similar in the first nine points; ferret and cat were similar to each other only in the last two points. There was no difference in retinal thickness among these three animals.     

亮叶杨桐(石芽茶)中黄酮类成分的研究

从广西产亮叶杨桐(Adinandra nitida Merr.ex H.L.Li)中,分得三种黄酮类成分:Ⅰ、Ⅱ和Ⅲ。经IR、UV(位移诊断)、熔点、混熔点、薄层层析鉴定:Ⅰ为芹菜素(Apigenin);Ⅱ为芹菜素-5-0-α-L-吡喃鼠李糖基(1→4-6~(?)-乙酰基-β-D-吡喃葡萄糖吡;Ⅲ为芹菜素-5-0-α-L-吡喃鼠李糖基(1→4)-β-D-吡喃葡萄糖甙。     

水蒸汽蒸馏巴柑檬叶和果皮精油化学成分的研究

巴柑檬是我们第一次从国外引种成功的一种名贵香料植物。用色谱-质谱-计算机联用技术、毛细管气相色谱保留指数法和标准品叠加法分析了水蒸汽蒸馏巴柑檬叶和果皮精油的化学成分。从叶和果皮精油分离出来的220个和200个成分中,分别鉴定出48个和57个成分。鉴定组分的含量分别占叶和果皮精油的99.80％和98.54％。叶精油与果皮精油在化学成分方面的主要区别是叶油中萜烃化合物含量较低,而萜醇类化合物含量较高。