Decreased methylation of the major mouse long interspersed repeated DNA during aging and in myeloma cells

Sequences of DNA that hybridize on Southern blots with cloned EcoR1 1.3 kb (ER1) of long interspersed repeated sequence (L1Md) of mouse have been examined in genomic DNA of neonatal mice, livers and brains of adult mice (3, 10, 27, and 30 mo old), and the solid myeloma tumor MOPC-315. The isoschizomers Hpa II (CCGG or mCCGG) and Msp I (CCGG or CmCGG) were used to assess methylation. We found that the L1Md sequence is fully methylated in young animals but demethylated in myeloma. Demethylation of L1Md sequence also occurred in aged animals. By scanning the autoradiogram, we found that approximately 8% of the 10(4)-10(5) copies have been demethylated in 27-mo-old liver.     

对TMV不同抗性番茄品种的叶绿体DNA限制性内切酶酶谱分析

选用对TMV有抗性和敏感的番茄品种、制备其ct-DNA, 用限制性内切酶BumHI、EcoRI和PstI完全酶解, 三种酶切图谱与前人报道一致, 由酶切片段计算番茄ct-DNA。分子量约为156.9kb。比较抗性和敏感品种的ct-DNA图谱, 发现三种酶切图谱均存在差异, 但由差异片段计算分子量之和又很除近。我们推测这是由于检基顺序变异或小段DNA顺序插入或缺失所造成, 由此证明, 叶绿体基因组与核中的TMV抗性基因, 共同决定着植物体对TMV的抗性。     

Targeting IFN-alpha to B cell lymphoma by a tumor-specific antibody elicits potent antitumor activities

IFN-alpha, a cytokine crucial for the innate immune response, also demonstrates antitumor activity. However, use of IFN-alpha as an anticancer drug is hampered by its short half-life and toxicity. One approach to improving IFN-alpha's therapeutic index is to increase its half-life and tumor localization by fusing it to a tumor-specific Ab. In the present study, we constructed a fusion protein consisting of anti-HER2/neu-IgG3 and IFN-alpha (anti-HER2/neu-IgG3-IFN-alpha) and investigated its effect on a murine B cell lymphoma, 38C13, expressing human HER2/neu. Anti-HER2/neu-IgG3-IFN-alpha exhibited potent inhibition of 38C13/HER2 tumor growth in vivo. Administration of three daily 1-microg doses of anti-HER2/neu-IgG3-IFN-alpha beginning 1 day after tumor challenge resulted in 88% of the mice remaining tumor free. Remarkably, anti-HER2/neu-IgG3-IFN-alpha demonstrated potent activity against established 38C13/HER2 tumors, with complete tumor remission observed in 38% of the mice treated with three daily doses of 5 microg of the fusion protein (p = 0.0001). Ab-mediated targeting of IFN-alpha induced growth arrest and apoptosis of lymphoma cells contributing to the antitumor effect. The fusion protein also had a longer in vivo half-life than rIFN-alpha. These results suggest that IFN-alpha Ab fusion proteins may be effective in the treatment of B cell lymphoma.     

Optimizing clinical dosing of combination broadly neutralizing antibodies for HIV prevention

Broadly neutralizing antibodies (bNAbs) are promising agents to prevent HIV infection and achieve HIV remission without antiretroviral therapy (ART). As with ART, bNAb combinations are likely needed to cover HIV’s extensive diversity. Not all bNAbs are identical in terms of their breadth, potency, and in vivo longevity (half-life). Given these differences, it is important to optimally select the composition, or dose ratio, of combination bNAb therapies for future clinical studies. We developed a model that synthesizes 1) pharmacokinetics, 2) potency against a wide HIV diversity, 3) interaction models for how drugs work together, and 4) correlates that translate in vitro potency to clinical protection. We found optimization requires drug-specific balances between potency, longevity, and interaction type. As an example, tradeoffs between longevity and potency are shown by comparing a combination therapy to a bi-specific antibody (a single protein merging both bNAbs) that takes the better potency but the worse longevity of the two components. Then, we illustrate a realistic dose ratio optimization of a triple combination of VRC07, 3BNC117, and 10–1074 bNAbs. We apply protection estimates derived from both a non-human primate (NHP) challenge study meta-analysis and the human antibody mediated prevention (AMP) trials. In both cases, we find a 2:1:1 dose emphasizing VRC07 is nearly optimal. Our approach can be immediately applied to optimize the next generation of combination antibody prevention and cure studies.     

Discovery and identification of anti-U1-A snRNP antibody in lung cancer

There are multiple reports of autoimmune response in patients with lung cancer. To investigate whether a novel autoantibody is present in patients with lung cancer and evaluate its clinical diagnostic and prognostic value, sera from 10 patients with lung cancer and 10 normal individuals were analyzed using immunofluorescence and Western blotting. It was found that one serum sample from the patients with squamous carcinoma gave a fine speckled pattern staining in nucleus and had a high titer antinuclear autoantibody which could recognize 31 kD of nuclear protein isolated from both cancer cells and normal cells. The same patient’s serum was further used to immunoprecipitate the target antigen. The protein bands were excised from the SDS-PAGE gels and were analyzed with a Qstar Pulser I Quadrupole time-flight mass spectrometer, and the 31 kD target antigen was identified as U1-AsnRNP. To test the prevalence of anti-U1-AsnRNP antibody, sera from 93 patients including 36 squmaous carcinomas (SCC), 26 adenocarcinomas (Ad), and 31 small cell carcinomas (SCLC) were screened by Western blotting. The results demonstrated that anti-U1-A snRNP antibody was present in 50% of SCC sera, 26.9% of Ad sera and 54.8% of SCLC sera. In this paper, we report for the first time that anti-U1-AsnRNP antibody could be detected in the patients with lung cancer.     

A highly selective and colorimetric assay of lysine by molecular-driven gold nanorods assembly

In this contribution, a simple, rapid, colorimeteric and selective assay for lysine was achieved by a controllable end-to-end assembly of gold nanorods (AuNRs) in the presence of Eu(3+) and lysine. This one-pot end-to-end assembly of 11-mercaptoundecanoic acid (MUA) modified AuNRs was occurred in Britton-Robinson buffer of pH 6.0, which involves the coordination binding between Eu(3+) and COO(-) groups as well as the electrostatic interaction of the COO(-) groups of MUA with the -NH(3)(+) group of lysine. As monitored by absorption spectra, scanning electron microscopic (SEM) images and dynamic light scattering (DLS) measurement, the end-to-end chain assembly results in large red-shift in the longitudinal plasmon resonance absorption (LPRA), giving red-to-blue color change of AuNRs. Importantly, it was found that the red-shift of LPRA is linearly proportional to the concentrations of lysine in the range of 5.0×10(-6)-1.0×10(-3)M with the limit of detection (LOD) being 1.6×10(-6)M (3σ/k). This red-shift of LPRA is highly selective, making it possible to develop a rapid, selective and visual assay for lysine in food samples.     

豚鼠冰水游泳应激后血浆、肾上腺髓质和脑内组织的亮—脑啡肽含量变化

豚鼠冰水游泳应激3min后,用放射免疫法测定血浆、肾上腺髓质和脑内三个脑区组织的亮—脑啡肽(Leu-enkephalin,LEK,)含量和血浆的血管紧张肽Ⅱ(Angiotensin Ⅱ,ATⅡ)浓度变化。结果表明:在冰水中游泳应激组豚鼠的血浆、肾上腺髓质、下丘脑和纹状体组织的LEK含量和血浆的ATⅡ浓度,均较对照组豚鼠明显升高(P值均<0.01)。提示:豚鼠在冰水中紧张、剧烈游泳后,可能激活了中枢神经和周围组织内的脑啡肽能神经元和血管紧张肽原酶—血管紧张肽Ⅱ系统,从而促进LEK和ATⅡ的释放增加。