Thermosensitive Nanoparticles with pH-Triggered Degradation and Release of Anti-inflammatory Cell-Penetrating Peptides

Poly(N-isopropylacrylamide-2-acrylamido-2-methyl-1-propanesulfonate) [poly(NIPAm-AMPS)] nanoparticles can be cross-linked with hydrolytically degradable N,O-dimethacryloyl hydroxylamine (DMHA) in order to yield a pH-sensitive drug delivery system that slowly erodes above pH 5.0. Varying the composition of degradable DMHA and nondegradable MBA cross-linking allows for engineered variation of particle size and degradation kinetics. Utilizing sulfated comonomer AMPS provides for increased passive loading of anti-inflammatory mitogen-activated protein kinase-activated protein kinase 2 (MK2)-inhibiting cell-penetrating peptide KAFAKLAARLYRKALARQLGVAA (KAFAK) between 24.3% and 29.2% (w/w) for nanoparticles with 5 mol % cross-linker. Nanoparticles were shown to be nontoxic in vitro and were effective at delivering a therapeutically active dose of KAFAK to THP1 human monocytes to suppress tumor necrosis factor α (TNF-α) expression during lipopolysaccharide (LPS)-induced inflammation. This thermosensitive nanoparticle system is an excellent platform for passive diffusive loading in deionized water and release in physiologically relevant ionic strength media of environmentally sensitive peptide therapeutics.     

医学院校新生适应性问题的研究

目的:在教育理论的指导下,通过问卷调查的形式对一年级学生的生活、学习、心理健康状况等方面的适应性问题进行调研。方法:采用随机抽样的原则,以抽到的班级所有符合条件的大学新生作为调查对象。采用自制的一般人口学特征问卷、高考志愿填报情况调查表、教学软硬件建设满意度调查表、目前学习状况及未来就业去向调查表、症状自评量表对46名大学新生进行调查。结果:调查班级学生中,多数学生来自农村家庭;与"70版、80后"相比,"90后"学生在职业选择方面具有更多的自主选择权;对学校在软硬条件方面亦提出更高的要求;受限于近期个人目标,缺乏长远人生目标,可持续发展能力差。心理问题阳性检出率为24%,排名在前4位的心理健康问题依次是:强迫、抑郁、焦虑、躯体化,男女生表现个有不同。结论:传统教育思维仍试图在极短时间里完成适应性教育工作,致使抽象理论与学生现实需求相脱节。陌生的环境、迥然不同的学习方式、全方位转变的人际关系使得新生心理问题出现叠加性、多重性的特点。相关部门应改变适应教育模式,建立新生适应教育活动的长期反馈机制和评估体系,才能更好的服务于新生教育工作。     

The microbiology of asthma

Asthma remains an important human disease that is responsible for substantial worldwide morbidity and mortality. The causes of asthma are multifactorial and include a complex mix of environmental, immunological and host genetic factors. In addition, epidemiological studies show strong associations between asthma and infection with respiratory pathogens, including common respiratory viruses such as rhinoviruses, human respiratory syncytial virus, adenoviruses, coronaviruses and influenza viruses, as well as bacteria (including atypical bacteria) and fungi. In this Review, we describe the many roles of microorganisms in the risk of developing asthma and in the pathogenesis of and protection against the disease, and we discuss the mechanisms by which infections affect the severity and prevalence of asthma.     

A Decade of Experience: Cryptococcus gattii in British Columbia

It has been over a decade since Cryptococcus gattii was first recognized as the causative organism of an outbreak of cryptococcosis on Vancouver Island, British Columbia. A number of novel observations have been associated with the study of this emergent pathogen. A novel genotype of C. gattii, VGIIa was described as the major genotype associated with clinical disease. Minor genotypes, VGIIb and VGI, are also responsible for disease in British Columbians, in both human and animal populations. The clinical major genotype VGIIa and minor genotype VGIIb are identical to C. gattii isolated from the environment of Vancouver Island. There is more heterogeneity in VGI, and a clear association with the environment is not apparent. Between 1999 and 2010, there have been 281 cases of C. gattii cryptococcosis. Risk factors for infection are reported to be age greater than 50 years, history of smoking, corticosteroid use, HIV infection, and history of cancer or chronic lung disease. The major C. gattii genotype VGIIa is as virulent in mice as the model Cryptococcus, H99 C. neoformans, although the outbreak strain produces a less protective inflammatory response in C57BL/6 mice. The minor genotype VGIIb is significantly less virulent in mouse models. Cryptococcus gattii is found associated with native trees and soil on Vancouver Island. Transiently positive isolations have been made from air and water. An ecological niche for this organism is associated within a limited biogeoclimatic zone characterized by daily average winter temperatures above freezing.     

The ATPase Activity of the P-glycoprotein Drug Pump Is Highly Activated When the N-terminal and Central Regions of the Nucleotide-binding Domains Are Linked Closely Together

The P-glycoprotein (P-gp, ABCB1) drug pump protects us from toxic compounds and confers multidrug resistance. Each of the homologous halves of P-gp is composed of a transmembrane domain (TMD) with 6 TM segments followed by a nucleotide-binding domain (NBD). The predicted drug- and ATP-binding sites reside at the interface between the TMDs and NBDs, respectively. Crystal structures and EM projection images suggest that the two halves of P-gp are separated by a central cavity that closes upon binding of nucleotide. Binding of drug substrates may induce further structural rearrangements because they stimulate ATPase activity. Here, we used disulfide cross-linking with short (8 Å) or long (22 Å) cross-linkers to identify domain-domain interactions that activate ATPase activity. It was found that cross-linking of cysteines that lie close to the LSGGQ (P517C) and Walker A (I1050C) sites of NBD1 and NBD2, respectively, as well as the cytoplasmic extensions of TM segments 3 (D177C or L175C) and 9 (N820C) with a short cross-linker activated ATPase activity over 10-fold. A pyrylium compound that inhibits ATPase activity blocked cross-linking at these sites. Cross-linking between the NBDs was not inhibited by tariquidar, a drug transport inhibitor that stimulates P-gp ATPase activity but is not transported. Cross-linking between extracellular cysteines (T333C/L975C) predicted to lock P-gp into a conformation that prevents close NBD association inhibited ATPase activity. The results suggest that trapping P-gp in a conformation in which the NBDs are closely associated likely mimics the structural rearrangements caused by binding of drug substrates that stimulate ATPase activity.     

Thoracic Aortic Aneurysm (TAAD)-causing Mutation in Actin Affects Formin Regulation of Polymerization

More than 30 mutations in ACTA2, which encodes α-smooth muscle actin, have been identified to cause autosomal dominant thoracic aortic aneurysm and dissection. The mutation R256H is of particular interest because it also causes patent ductus arteriosus and moyamoya disease. R256H is one of the more prevalent mutations and, based on its molecular location near the strand-strand interface in the actin filament, may affect F-actin stability. To understand the molecular ramifications of the R256H mutation, we generated Saccharomyces cerevisiae yeast cells expressing only R256H yeast actin as a model system. These cells displayed abnormal cytoskeletal morphology and increased sensitivity to latrunculin A. After cable disassembly induced by transient exposure to latrunculin A, mutant cells were delayed in reestablishing the actin cytoskeleton. In vitro, mutant actin exhibited a higher than normal critical concentration and a delayed nucleation. Consequently, we investigated regulation of mutant actin by formin, a potent facilitator of nucleation and a protein needed for normal vascular smooth muscle cell development. Mutant actin polymerization was inhibited by the FH1-FH2 fragment of the yeast formin, Bni1. This fragment strongly capped the filament rather than facilitating polymerization. Interestingly, phalloidin or the presence of wild type actin reversed the strong capping behavior of Bni1. Together, the data suggest that the R256H actin mutation alters filament conformation resulting in filament instability and misregulation by formin. These biochemical effects may contribute to abnormal histology identified in diseased arterial samples from affected patients.