Enhanced cytotoxicity of saporin by polyamidoamine dendrimer conjugation and photochemical internalization

Because of the lack of membrane binding subunit, type I ribosome-inactivating proteins (RIPs) are not very toxic to cells unless action is taken to allow for toxin internalization to the cytosol. To overcome the potential barriers that greatly hinder the cellular uptake and intracellular release of saporin, a type I RIP, we used generation 4 polyamidoamine (PAMAM) dendrimer as the carrier to improve its endocytic uptake, passive tumor targeting, and implemented the photochemical internalization (PCI) technology to facilitate its cytosolic release. Our results showed that the cellular uptake of saporin was increased after conjugation with the PAMAM dendrimer and the cytotoxic effect was improved by more than 1 order of magnitude. The cytotoxicity of free saporin and PAMAM-saporin was further enhanced by the PCI technology. PCI changed the mechanism of cellular uptake of free saporin and then caused more saporin entering into the cells. After the PCI treatment, PAMAM-saporin was not only internalized into the cytosol but also efficiently entered the nuclei. Our results indicated that conjugating to PAMAM dendrimer is a possible approach to enhance the cellular uptake of saporin. PCI is a promising technology to significantly enhance the cytotoxicity of both free saporin and PAMAM-saporin. Combining both polymer conjugation and PCI approaches may improve the efficacy of RIPs in cancer therapy.     

Pivotal advance: HMGB1 expression in active lesions of human and experimental multiple sclerosis

Multiple sclerosis (MS) is a chronic, inflammatory, demyelinating disease of the CNS, most frequently starting with a series of bouts, each followed by complete remission and then a secondary, progressive phase during which the neurological deficit increases steadily. The underlying molecular mechanisms responsible for disease progression are still unclear. Herein, we demonstrate that high mobility group box chromosomal protein 1 (HMGB1), a DNA-binding protein with proinflammatory properties, is evident in active lesions of MS and experimental autoimmune encephalomyelitis (EAE) and that HMGB1 levels correlate with active inflammation. Furthermore, the expression of the innate HMGB1 receptors--receptor for advanced glycation end products, TLR2, and TLR4--was also highly increased in MS and rodent EAE. Additionally, in vitro activation of rodent CNS-derived microglia and bone marrow-derived macrophages demonstrated that microglia were equally as capable as macrophages of translocating HMGB1 following LPS/IFN-gamma stimulation. Significant expression of HMGB1 and its receptors on accumulating activated macrophages and resident microglia may thus provide a positive feedback loop that amplifies the inflammatory response during MS and EAE pathogenesis.     

Safety and efficacy of gene transfer for Leber's congenital amaurosis

Leber's congenital amaurosis (LCA) is a group of inherited blinding diseases with onset during childhood. One form of the disease, LCA2, is caused by mutations in the retinal pigment epithelium-specific 65-kDa protein gene (RPE65). We investigated the safety of subretinal delivery of a recombinant adeno-associated virus (AAV) carrying RPE65 complementary DNA (cDNA) (ClinicalTrials.gov number, NCT00516477 [ClinicalTrials.gov]). Three patients with LCA2 had an acceptable local and systemic adverse-event profile after delivery of AAV2.hRPE65v2. Each patient had a modest improvement in measures of retinal function on subjective tests of visual acuity. In one patient, an asymptomatic macular hole developed, and although the occurrence was considered to be an adverse event, the patient had some return of retinal function. Although the follow-up was very short and normal vision was not achieved, this study provides the basis for further gene therapy studies in patients with LCA.     

Homozygous bisalbuminemia or homozygous mutant albumin?

IntroductionBisalbuminemia is a genetic condition in which an albumin variant is found in serum in addition to normal albumin.Methods and materialsSerum protein electrophoresis using the Sebia HYDRASYS electrophoresis system was performed on an 84 year old male.ResultsSerum protein electrophoresis showed a single albumin band migrating faster than normal albumin.ConclusionThe presence of a homozygous albumin variant band, albumin Naskapi, is noted.     

A cross-sectional assessment of stroke rehabilitation in nebraska hospitals

Jones KJ, Cochran TM, Jensen LE, Roehrs TG, Volkman KG, Goldman AJ. A cross-sectional assessment of stroke rehabilitation in Nebraska hospitals.ObjectiveTo assess the structure and process of stroke rehabilitation in Nebraska hospitals.DesignCross-sectional mail survey using the Dillman tailored-design method of administration.SettingHospitals in Nebraska.ParticipantsApproximately 77% of the 84 Nebraska hospitals that provide stroke rehabilitation are critical access hospitals (CAHs) that are limited to 25 beds. Our study sample of hospitals (N=53) included the 19 hospitals licensed for 47 to 689 beds (non-CAHs) and a stratified random sample of 34 of the 65 CAHs.InterventionsNot applicable.Main Outcome MeasuresSelf-reported stroke rehabilitation team structure and processes, purposes of and barriers to the use of evidence-based standardized assessments, specific assessments used, and access to specialized stroke rehabilitation services and community resources.ResultsThirty-six (68%) of the 53 hospitals responded to the survey. Approximately 61% of the hospitals used an organized team to provide stroke rehabilitation; 8% of the hospitals—all non-CAHs—had a team dedicated to stroke rehabilitation. After adjusting for hospital size, having an organized team was significantly associated with the use of standardized assessments to improve communication, measure progress and outcomes, evaluate effectiveness of practice, and compare patient outcomes across conditions. Access to specialized stroke rehabilitation professionals and services was significantly greater in non-CAHs.ConclusionsHospital size and the presence of a team are determinants of the structure and process of stroke rehabilitation in Nebraska hospitals. Further research is needed to determine (1) whether team structure is a determinant of stroke rehabilitation outcomes across the continuum of care settings, (2) the needs of rural stroke survivors, and (3) whether technology can facilitate the use of stroke rehabilitation standardized assessments by rural health care professionals.     

Changes in Peripheral CD4+CD25high Regulatory T Cells in the Acute-on-Chronic Liver Failure Patients with Plasma Exchange Treatment

The prevalence of CD4+CD25^high regulatory T cells (Tregs) in patients with acute-on-chronic liver failure (AoCLF) who received plasma exchange (PE) and/or medical treatment was investigated. One hundred five patients with AoCLF in two groups (PE plus routine-care, n = 48 and routine-care, n = 57) were enrolled in our study. In the PE group, there were 27 survivors (27/48) while, in the routine-care group, there were 18 survivors (18/57), both after 30 days treatment. Twenty-three healthy donors were used as the control group. Tregs were determined by flow cytometry serially. In the survivors, Tregs frequency were lower compared with the normal controls on admission and showed an up and down tendency; moreover, this frequency turned to the level as that in healthy subjects and was faster in the PE compared with the medical group while, among the nonsurvivors, Tregs stayed at a high level throughout the examination period. Importantly, an increased quantity of Tregs was associated with high mortality and reduced survival time of AoCLF patients. These data suggest that Tregs play a role in determining the patient’s fate toward either a favorable or unfavorable clinical course of disease, and PE may represent a reliable hepatic support device for AoCLF.     

The Transcription Levels of ABCA1, ABCG1 and SR-BI are Negatively Associated with Plasma CRP in Chinese Populations with Various Risk Factors for Atherosclerosis

ATP binding cassette transporters (ABCA1, ABCG1) and scavenger receptor class B type I (SR-BI) are the three most important cellular cholesterol transporters that may prevent atherogenesis. The aim of this study was to investigate whether they were altered in Chinese populations with various risk factors for atherosclerosis and their potential associations with C-reactive protein (CRP). Healthy female controls (n?=?30) and populations with various risk factors for atherosclerosis, such as type 2 diabetes (n?=?17), hypertension (n?=?12), overweight/obesity (n?=?10), incipient nephropathy (n?=?10), postmenopausal women (n?=?9), male (n?=?19), ageing male (n?=?22), or smoking (n?=?16), were recruited. ABCA1, ABCG1 and SR-BI mRNA levels in peripheral monocytes was determined. ABCG1 was decreased in all the risk populations except ageing. ABCA1 was decreased in all the risk populations except diabetes and male. SR-BI was decreased in those with overweight/obesity and incipient nephropathy. Circulating CRP was increased almost in all the risk populations except in males. The levels of ABCA1, ABCG1 and SR-BI were reduced in those with subclinically high CRP, and negatively associated with CRP level. These data indicates that ABCA1, ABCG1, and SR-BI are reduced in various populations under subclinically inflammatory conditions, which may potentially lead to impairing reverse cholesterol transport and developing atherosclerosis.     

高龄急性心肌梗死患者经皮冠状动脉介入治疗的临床分析

目的探讨经皮冠状动脉介入治疗(PCI)高龄急性心肌梗死(AMI)患者临床特点、影响预后的相关因素。方法将AMI患者176例分为高龄组(≥75岁,74例)和非高龄组(<75岁,102例);对比分析病史、左室舒张末内径(LVDD)和左室射血分数(LVEF),检测PCI术前血清肌酸激酶同工酶(CK-MB)、血清肌钙蛋白I(cTnI)、高敏C-反应蛋白(hsCRP)和血肌酐(Cr),分析两组患者的性别、危险因素、并发症等;观察PCI特点及术后主要不良心血管事件(MACE)发生情况。结果与非高龄组比较,高龄组合并高血压、2型糖尿病、缺血性脑卒中及慢性阻塞性肺疾病的发生率明显增高,LVDD增大,LVEF降低,CK-MB、hsCRP、cTnI、Cr水平显著增高,病变血管数和复杂病变数及术后并发症均明显增多,且住院时间长,在住院及随访12个月期间MACE发生率明显增高。结论高龄AMI患者合并多种慢性疾病,冠状动脉病变复杂,PCI并发症多,预后较差。