The STING activator c-di-AMP exerts superior adjuvant properties than the formulation poly(I:C)/CpG after subcutaneous vaccination with soluble protein antigen or DEC-205-mediated antigen targeting to dendritic cells

Vaccination is the most efficient strategy to protect from infectious diseases and the induction of a protective immune response not only depends on the nature of the antigen, but is also influenced by the vaccination strategy and the co-administration of adjuvants. Therefore, the precise monitoring of adjuvant candidates and their immune modulatory properties is a crucial step in vaccine development. Here, one central aspect is the induction of appropriate humoral and cellular effector mechanisms.In our study we performed a direct comparison of two promising candidates in adjuvant development, the STING activator bis-(3,5)-cyclic dimeric adenosine monophosphate (c-di-AMP) and the Toll-like receptor ligand formulation poly(I:C)/CpG. These were evaluated in C57BL/6 mice using the model antigen ovalbumin (OVA) in subcutaneous vaccination with soluble protein as well as in a dendritic cell (DC) targeting approach (αDEC-OVA). Strikingly, c-di-AMP as compared to poly(I:C)/CpG resulted in significantly higher antigen-specific IgG antibody levels when used in immunization with soluble OVA as well as in antigen targeting to DC. In vaccination with soluble OVA, c-di-AMP induced a significantly stronger CTL, Th1 and IFNγ-producing CD8⁺ memory T cell response than poly(I:C)/CpG. The response was CTL and Th1 cell dominated, a profile shared by both adjuvants. In the context of targeting OVA to DC, c-di-AMP induced significantly increased Th1 and Th2 cell responses as compared to poly(I:C)/CpG. Interestingly, the Th1 response dominated the overall T cell response only when c-di-AMP was used, indicating a distinct modulatory property of c-di-AMP when the DC targeting immunization approach was exploited.Taken together, we describe superior properties of c-di-AMP as compared to poly(I:C)/CpG in subcutaneous vaccination with soluble antigen as well as antigen targeting to DC. This indicates exceptionally effective adjuvant properties for c-di-AMP and provides compelling evidence of its potential for further adjuvant development, especially also when using DC targeting approaches.     

三子颗粒下调miR-205-5p抑制小鼠脾虚型肠道腺瘤生长研究

目的 探讨三子颗粒通过降低微小核糖核酸-205-5p（miR-205-5p）水平抑制小鼠脾虚型肠道腺瘤生长的作用。方法 取70只4周龄的雄性C57BL/6J小鼠,采用对氧化偶氮甲烷（AOM）/葡聚糖硫酸钠（DSS）诱导小鼠结直肠腺瘤模型,将建模小鼠随机分为模型组、阿司匹林（200 mg·kg^-1）组、miR inhibitor-NC （2 mg·kg^-1）组、miR-205-5p inhibitor （2 mg·kg^-1）组和三子颗粒低、中、高剂量（1.7、3.4、6.8 g·kg^-1）组,造模期间ig给药,每天1次。比较各组小鼠肠道腺瘤的数量并测量腺瘤体积;苏木素-伊红（HE）染色观察小鼠肠道腺瘤的病理情况;CCK-8法检测各组小鼠肠道腺瘤细胞增殖活力;原位末端标记（TUNEL）法检测小鼠肠道腺瘤细胞凋亡;实时荧光定量PCR（qRT-PCR）法检测肠道腺瘤miR-205-5p表达量及磷酸酯酶与张力蛋白同源物（PTEN）、B淋巴细胞瘤-2（Bcl-2）、Bcl-2相关X蛋白（Bax）、Ki67 mRNA表达量;Western blotting法检测PTEN、Bcl-2、Bax、Ki67蛋白表达水平;荧光素酶活性实验验证miR-205-5p和PTEN的靶向关系。结果 与模型组比较,阿司匹林组和三子颗粒低、中、高剂量组小鼠肠道腺瘤数量、体积及细胞增殖活性均显著降低,凋亡率显著升高（P＜0.05）,miR-205-5p表达量、Bcl-2、Ki67 mRNA及蛋白表达量显著降低,PTEN、Bax mRNA及蛋白表达量显著升高（P＜0.05）,其中三子颗粒作用呈剂量相关性;与miR inhibitor-NC组比较,miR-205-5p inhibitor组小鼠肠道腺瘤数量、体积及细胞增殖活性均显著降低,凋亡率显著升高（P＜0.05）,miR-205-5p表达量、Bcl-2、Ki67 mRNA及蛋白表达量显著降低,PTEN、Bax mRNA及蛋白表达量显著升高（P＜0.05）;荧光素酶活性实验证实miR-205-5p可靶向调控PTEN。结论 三子颗粒可抑制小鼠脾虚型肠道腺瘤生长,可能是通过下调miR-205-5p,上调PTEN、Bax表达,下调Bcl-2、Ki67表达发挥作用的。     

Elimination of Lymphatic Filariasis: Mass Drug Administration in Endemic Areas of (Bidar District) Karnataka-2008

Background:

Lymphatic Filariasis is a mosquito transmitted disease, caused by parasitic worm Wuchereria bancrofti. Global Programme for Elimination of Lymphatic Filariasis was established in early 2000. The strategy recommended by the World Health Organization is annual Mass Drug Administration (MDA) of single-dose of Diethylcarbamazine 6 mg/kg (DEC), distributed to inhabitants of Filariasis endemic areas, excluding children below 2 years of age, pregnant women, and seriously ill persons, and Morbidity Management. The health system distributes the drugs by a door-to-door strategy.

Objective:

To assess the coverage and compliance of MDA in Bidar district during the campaign in November 2008.

Materials and Methods:

Cross-sectional population-based house-to-house visit. Outcome is assessed as actual coverage and compliance, in Percentage and proportions.

Results:

Eight clusters, total eligible population of 1 131 individuals were interviewed. The coverage rate was 78% with variation across different areas. The compliance with drug ingestion was 68%.

Conclusion:

The effective coverage was below the target (85%). Side effects of DEC were minimum, the overall coverage was better in rural areas compared with urban areas.     

A simple approach for enhanced immune response using engineered dendritic cell targeted nanoparticles

In this study, we demonstrate a simple strategy for enhanced immune response using a two-component dendritic cell (DC) targeted antigen delivery system. One component consists of a recombinant bifunctional fusion protein (bfFp) used for DC targeting, whereas, the other component is made of biotinylated PLGA nanoparticles that encapsulate the antigen. The fusion protein (bfFp) made of a truncated core-streptavidin fused to anti-DEC-205 single chain antibody (scFv) was mixed with ovalbumin-loaded biotinylated NPs that were formulated using biotin–PEG (2000)–PLGA, and the combination, bfFp functionalized NPs was used for DC targeted antigen delivery. In vitro DC uptake studies revealed a 2-fold higher receptor-mediated uptake of bfFp functionalized NPs when compared to non-targeted NPs. Immunization of the mice with the bfFp functionalized NPs in conjunction with DC maturation stimulus (anti-CD40 mAb) enhanced OVA-specific IgG and IgG subclass responses. Splenocytes of these mice secreted significantly higher levels of Th1 (IFN-γ and IL-2) cytokines upon ex vivo restimulation with OVA. The promising outcomes of the bfFp functionalized DC targeted system support its use as a versatile vaccine delivery system for the design of monovalent or polyvalent vaccines.     

Capillary electrophoresis–mass spectrometry: Recent trends in clinical proteomics

The increasing attention now paid to the elucidation of human proteome strengthened the development of analytical instruments able to provide reliable proteins and peptides quantitation and characterization in biological fluids and tissues. Emerging from proteomics, clinical proteomics exclusively considers its biomedical applications. It evaluates, often by high-throughput comparative platforms, the protein and peptide variations in body fluids, cells and tissues under different physiological and pathological conditions with the aim of discovering disease biomarkers. Among the available analytical methodologies, mass spectrometry in coupling with liquid chromatography or capillary electrophoresis demonstrated to be the eligible technique for protein detection and identification. This review summarizes the most recent applications of capillary electrophoresis–mass spectrometry to clinical proteomics, focusing on capillary zone electrophoresis separation mode and ESI and MALDI ionizations, which are the most frequently applied capillary electrophoresis–mass spectrometry hyphenated techniques.     

结肠癌细胞系Colo205细胞膜表面表达血小板和T细胞活化抗原1的配体

目的　对血小板和T细胞活化抗原 1(PTA1)配体的表达做初步鉴定。方法　首先克隆PTA1胞膜外区基因片段、构建PTA1/Ig融合蛋白表达载体 ,制备PTA1/Ig融合蛋白。通过粘附实验及组化实验证实PTA1配体的分布。结果　PTA1/Ig能特异性结合于Colo2 0 5细胞表面 ,并能阻断活化Jurkat细胞对Colo2 0 5细胞的粘附。结论　证实结肠癌细胞系Colo2 0 5表达PTA1配体 ,为今后进一步了解肿瘤细胞与活化T细胞的相互作用在肿瘤发病机理中的意义创造了条件。     

Dynamic mislocalizations of nuclear pore complex proteins after focal cerebral ischemia in rat

Nuclear pore complexes (NPCs) play an important role in coordinating the transport of proteins and nucleic acids between the nucleus and cytoplasm, and are therefore essential for maintaining normal cellular function and liability. In the present study, we investigated the temporal immunohistochemical distribution of five representative components of NPCs—Ran GTPase‐activating protein 1 (RanGap1), glycoprotein‐210 (Gp210), nucleoporin 205 (Nup205), nucleoporin 107 (Nup107), and nucleoporin 50 (Nup50)—after 90 min of transient middle cerebral artery occlusion (tMCAO) up to 28 days after the reperfusion in rat brains. Single immunohistochemical analyses showed ring‐like stainings along the periphery of the nucleus in sham control brains. After tMCAO, Gp210 and Nup107 immunoreactivity continuously increased from 1 day, and RanGap1, Nup205, and Nup50 increased from 2 days until 28 days, which also displayed progressive precipitations within the nucleus in the peri‐ischemic area, while the ischemic core showed scarce expression with collapsed structure. Double immunofluorescent analyses revealed nuclear retention and apparent colocalization of RanGap1 with Nup205, Gp210 with Nup205, and partial colocalization of Nup205 with Nup107; most of the ischemic changes above were similar to those observed in patients with C9orf72‐genetic amyotrophic lateral sclerosis. Taken together, these observations suggest that the mislocalization of these nucleoporins may be a common pathogenesis of both ischemic and neurodegenerative disease. © 2016 Wiley Periodicals, Inc.     

Sleep and weight-height development

Objectives

To describe the association between sleep duration and weight–height development in children and adolescents.