The synthesis and SAR study of phenylalanine-derived (Z)-5-arylmethylidene rhodanines as anti-methicillin-resistant Staphylococcus aureus (MRSA) compounds

A focused library of rhodanine compounds containing novel substituents at the C5-position was synthesized and tested in vitro against a panel of clinically relevant MRSA strains. The present SAR study was based on our lead compound 1 (MIC = 1.95 μg/mL), with a focus on identifying optimal C5-arylidene substituents. In order to obtain this objective, we condensed several unique aromatic aldehydes with phenylalanine-derived rhodanine intermediates to obtain C5-substituted target rhodanine compounds for evaluation as anti-MRSA compounds. These efforts produced three compounds with significant efficacy: 23, 32 and 44, with MIC values ranging from 0.98 to 1.95 μg/mL against all tested MRSA strains as compared to the reference antibiotics penicillin G (MIC = 15.60–250.0 μg/mL) and ciprofloxacin (MIC = 7.80–62.50 μg/mL) and comparable to that of vancomycin (MIC = 0.48 μg/mL). In addition, compounds 24, 28, 37, 41, 46 and 48 (MIC = 1.95–3.90 μg/mL) were efficacious against all MRSA strains. The majority of the synthesized compounds had bactericidal activity at concentrations only two to fourfold higher than their MIC. Overall, the results suggest that compounds 23, 32 and 44 may be of potential use in the treatment of MRSA infections.     

Interaction of serum amyloid A with human cystatin C—assessment of amino acid residues crucial for hCC–SAA formation (part II)

Secondary amyloid A (AA) amyloidosis is an important complication of some chronic inflammatory diseases, primarily rheumatoid arthritis (RA). It is a serious, potentially life‐threatening disorder caused by the deposition of AA fibrils, which are derived from the circulatory, acute‐phase‐reactant, serum amyloid A protein (SAA). Recently, a specific interaction between SAA and the ubiquitous inhibitor of cysteine proteases—human cystatin C (hCC)—has been proved. Using a combination of selective proteolytic excision and high‐resolution mass spectrometry, the binding sites in the SAA and hCC sequences were assessed as SAA(86–104) and hCC(96–102), respectively. Here, we report further details concerning the hCC–SAA interaction. With the use of affinity tests and florescent ELISA‐like assays, the amino acid residues crucial for the protein interaction were determined. It was shown that all amino acid residues in the SAA sequence, essential for the formation of the protein complex, are basic ones, which suggests an electrostatic interaction character. The idea is corroborated by the fact that the most important residues in the hCC sequence are Ser‐98 and Tyr‐102; these residues are able to form hydrogen bonds via their hydroxyl groups. The molecular details of hCC–SAA complex formation might be helpful for the design of new compounds modulating the biological role of both proteins. Copyright © 2013 John Wiley & Sons, Ltd.     

小柴胡汤加减联合奥美拉唑对慢性胃炎具有治疗增效作用

本研究选取本院收治的慢性胃炎患者114例并根据治疗方法不同分为对照组和观察组。对照组患者采用奥美拉唑治疗,观察组患者采用奥美拉唑联合小柴胡汤加减治疗,分析小柴胡汤加减联合奥美拉唑对慢性胃炎血清表皮生长因子(EGF)、胃黏膜氧化酶-2 (COX-2)蛋白表达情况的影响及护理对策。研究结果表明,治疗前两组患者胃镜检查充血水肿、糜烂、黏膜白相、颗粒增生发生率比较无统计学差异(p>0.05);治疗后,观察组患者各项指标发生率低于对照组(p<0.05);治疗前,两组患者血清EGF、Bcl-2、CRP和胃黏膜COX-2、P-p65表达水平比较无统计学差异(p>0.05),治疗后发现观察组患者血清EGF水平高于对照组,血清Bcl-2、CRP和胃黏膜COX-2、P-p65表达水平低于对照组(p<0.05);治疗前,两组患者在躯体功能、躯体职能、躯体疼痛、情感职能、心理健康评分等生活质量评分上比较无统计学差异(p<0.05),治疗后,观察组患者各项评分高于对照组(p>0.05)。本研究初步结论说明,小柴胡汤加减联合奥美拉唑治疗慢性胃炎疗效显著,可改善患者血清EGF、Bcl-2和胃黏膜COX-2表达水平,提高患者生活质量。     

皮下种植黑色素瘤对小鼠血液中脂肪和氨基酸代谢的影响*

目的:探讨皮下种植黑色素瘤对小鼠血液中相关脂肪和氨基酸代谢的影响。方法:野生型小鼠皮下种植B16黑色素瘤细胞,两周后观察小鼠并取不同的组织称重,检测血液中甘油三酯、葡萄糖、脂肪酸、炎症因子的水平。利用质谱方法检测血液中氨基酸的水平。结果:野生型小鼠皮下肿瘤种植2周后体重没有明显变化,脾脏和肝脏的重量明显增加,皮下脂肪组织和腹腔脂肪组织的重量没有明显变化。血液中甘油三酯的水平明显增高,葡萄糖和甘油的水平明显下降。血液中炎症因子Interleukin 6(IL6)和Interferon gamma(IFNγ)的水平明显增加。血液的质谱结果显示有四种氨基酸的水平有明显变化,其中胱氨酸(Cystine)增加了约30倍,丝氨酸(Serine)、谷氨酰胺(Glutamine)和色氨酸(Tryptophan)分别有不同程度的降低。结论:皮下黑色素瘤显著影响小鼠机体营养状态和炎症,并特异性影响血液中氨基酸的代谢。     

Serum tau fragments as predictors of death or poor neurological outcome after out-of-hospital cardiac arrest

Background: Anoxic brain injury is the primary cause of death after resuscitation from out-of-hospital cardiac arrest (OHCA) and prognostication is challenging. The aim of this study was to evaluate the potential of two fragments of tau as serum biomarkers for neurological outcome.

Methods: Single-center sub-study of 171 patients included in the Target Temperature Management (TTM) Trial randomly assigned to TTM at 33?°C or TTM at 36?°C for 24?h after OHCA. Fragments (tau-A and tau-C) of the neuronal protein tau were measured in serum 24, 48 and 72?h after OHCA. The primary endpoint was neurological outcome.

Results: Median (quartile 1 – quartile 3) tau-A (ng/ml) values were 58 (43–71) versus 51 (43–67), 72 (57–84) versus 71 (59–82) and 76 (61–92) versus 75 (64–89) for good versus unfavourable outcome at 24, 48 and 72?h, respectively (p_group = 0.95). Median tau C (ng/ml) values were 38 (29–50) versus 36 (29–49), 49 (38–58) versus 48 (33–59) and 48 (39–59) versus 48 (36–62) (p_group = 0.95). Tau-A and tau-C did not predict neurological outcome (area under the receiver-operating curve at 48?h; tau-A: 0.51 and tau-C: 0.51).

Conclusions: Serum levels of tau fragments were unable to predict neurological outcome after OHCA.     

Suction blister fluid as potential body fluid for biomarker proteins

Early diagnosis is important for effective disease management. Measurement of biomarkers present at the local level of the skin could be advantageous in facilitating the diagnostic process. The analysis of the proteome of suction blister fluid, representative for the interstitial fluid of the skin, is therefore a desirable first step in the search for potential biomarkers involved in biological pathways of particular diseases. Here, we describe a global analysis of the suction blister fluid proteome as potential body fluid for biomarker proteins. The suction blister fluid proteome was compared with a serum proteome analyzed using identical protocols. By using stringent criteria allowing less than 1% false positive identifications, we were able to detect, using identical experimental conditions and amount of starting material, 401 proteins in suction blister fluid and 240 proteins in serum. As a major result of our analysis we construct a prejudiced list of 34 proteins, relatively highly and uniquely detected in suction blister fluid as compared to serum, with established and putative characteristics as biomarkers. We conclude that suction blister fluid might potentially serve as a good alternative biomarker body fluid for diseases that involve the skin.     

Expression of inflammatory cytokines in mesenchymal stromal cells is sensitive to culture conditions and simple cell manipulations

Background

Mesenchymal stromal cells (MSCs) can be used in several clinical applications. While MSCs are frequently cultured in fetal bovine serum for in vitro experimentation, human serum supplements are required for cells to be used in patients. Here we show how different human serum supplements and in vitro manipulations used during the cell culture impact on MSC proliferation rate and expression of inflammatory molecules.

低聚异麦芽糖对模拟失重大鼠肠道益生菌以及钙代谢和骨矿盐密度影响的初步研究

目的：研究低聚异麦芽糖对模拟失重大鼠肠道益生菌的影响以及与骨钙代谢变化的关系。方法：30只雄性SD大鼠随机分为3组（每组10只）FC组;普通对照组（饲喂普通饲料）;SC组,模拟失重对照组（饲喂普通饲料）;SS组,模拟失重低聚异麦芽糖组（饲喂普通饲料 低聚异麦芽糖）,实验期21d。结果：SC组大鼠肠道益生菌（主要是双歧杆菌和乳杆菌）数量、饲料钙的表观吸收率、股骨骨密度（BMD）,骨钙含量以及骨钙素（BGP）的水平显著低于FC组,而血钙水平明显高于FC组;SS组大鼠肠道益生菌数量,饲料钙的表观吸收率,骨密度,骨钙含量以及骨钙素水平较SC组高,血钙水平显著低于SC组。结论：低聚异麦芽糖可以促进模拟失重大鼠肠道益生菌的增殖,减少股骨骨质的丢失,提高股骨BMD,增加骨形成,对骨代谢产生一定的有益影响。     

Cell Cycle Arrest of Proliferating Neuronal Cells by Serum Deprivation Can Result in Either Apoptosis or Differentiation

Abstract: Apoptotic cell death plays a critical role in the development of the nervous system. The death of mature nondividing neurons that fail to receive appropriate input from the target field has been extensively studied. However, the mechanisms mediating the extensive cell death occurring in areas of the developing brain where proliferating neuroblasts differentiate into mature nondividing neurons have not been analyzed. We show here that the cell cycle arrest of a proliferating cell of neuronal origin by removal of serum results in either apoptotic cell death or differentiation to a mature nondividing neuronal cell. The proportion of cells undergoing death or differentiation is influenced in opposite directions by treatment of the cells with cyclic AMP and retinoic acid. This suggests that following the withdrawal of signals stimulating neuroblast cell division, neuronal cells either can cease to suppress a constitutive suicide pathway and hence die by apoptosis or, alternatively, can differentiate into a mature neuronal cell. Regulation of the balance between apoptosis and neuronal differentiation could therefore play a critical role in controlling the numbers of mature neurons that form.     

Activation of formyl peptide receptor like-1 by serum amyloid A induces CCL2 production in human umbilical vein endothelial cells

We investigated the effects of serum amyloid A (SAA) on the production of C-C chemokine motif ligand 2 (CCL2) and the mechanism underlying SAA action in human umbilical vein endothelial cells (HUVECs). Stimulation of HUVECs by SAA elicited CCL2 production in a concentration-dependent manner. SAA induced the activations of NF-κB and AP-1, which were essential for CCL2 production after SAA stimulation. HUVECs expressed formyl peptide receptor-like 1 (FPRL1), and short interfering RNA knockdown of FPRL1 nearly completely blocked SAA-induced CCL2 production in HUVECs. We suggest that SAA stimulates CCL2 production via FPRL1 and, thus, contributes to atherosclerosis.