A multiscale mathematical model of avascular tumor growth to investigate the therapeutic benefit of anti-invasive agents

With the aim of inhibiting cancer growth and reducing the risk of metastasis, pharmaceutical companies in the early 1990s developed anti-metastatic agents called inhibitors of metalloproteinases (MMPi). Despite the promising results obtained in pre-clinical studies, results of Phase III trials have been somewhat disappointing for late stage cancer patients. With the aim of mathematically investigating this therapeutic failure, we developed a mechanistically based model which integrates cell cycle regulation and macroscopic tumor dynamics. By simulating the model, we evaluated the efficacy of MMPi therapy. Simulation results predict the lack of efficacy of MMPi in advanced cancer patients. The theoretical model may aid in evaluating the efficacy of anti-metastatic therapies, thus benefiting the design of prospective clinical trials.     

Intercellular communication that mediates formation of the neuromuscular junction

Reciprocal signals between the motor axon and myofiber induce structural and functional differentiation in the developing neuromuscular junction (NMJ). Elevation of presynaptic acetylcholine (ACh) release on nerve-muscle contact and the correlated increase in axonal-free calcium are triggered by unidentified membrane molecules. Restriction of axon growth to the developing NMJ and formation of active zones for ACh release in the presynaptic terminal may be induced by molecules in the synaptic basal lamina, such as S-laminin, heparin binding growth factors, and agrin. Acetylcholine receptor (AChR) synthesis by muscle cells may be increased by calcitonin gene-related peptide (CGRP), ascorbic acid, and AChR-inducing activity (ARIA)/heregulin, which is the best-established regulator. Heparin binding growth factors, proteases, adhesion molecules, and agrin all may be involved in the induction of AChR redistribution to form postsynaptic-like aggregates. However, the strongest case has been made for agrin's involvement. “Knockout” experiments have implicated agrin as a primary anterograde signal for postsynaptic differentiation and muscle-specific kinase (MuSK), as a putative agrin receptor. It is likely that both presynaptic and postsynaptic differentiation are induced by multiple molecular signals. Future research should reveal the physiological roles of different molecules, their interactions, and the identity of other molecular participants.     

Deacylated lipopolysaccharides inhibit biofilm formation by Gram-negative bacteria

The extracellular polysaccharides of Vibrio vulnificus play different roles during biofilm development. Among them, the effect of lipopolysaccharide (LPS), which is crucial for bacterial adherence to surfaces during the initial stage of biofilm formation, on the formation process was examined using various types of LPS extracts. Exogenously added LPS strongly inhibited biofilm formation in a dose-dependent manner. In addition, the exogenous addition of a deacylated form of LPS (dLPS) also inhibited biofilm formation. However, an LPS fraction extracted from a mutant not able to produce O-antigen polysaccharides (O-Ag) did not have an inhibitory effect. Furthermore, biofilm formation by several Gram-negative bacteria was inhibited by dLPS addition. In contrast, biofilm formation by Gram-positive bacteria was not influenced by dLPS but was affected by lipoteichoic acid. Therefore, this study demonstrates that O-Ag in LPS is important for inhibiting biofilm formation and may serve an efficient anti-biofilm agent specific for Gram-negative bacteria.     

A dual approach in the study of poly (ADP-ribose) polymerase: In vitro random mutagenesis and generation of deficient mice

A dual approach to the study of poly (ADP-ribose)polymerase (PARP) in terms of its structure and function has been developed in our laboratory. Random mutagenesis of the DNA binding domain and catalytic domain of the human PARP, has allowed us to identify residues that are crucial for its enzymatic activity.In parallel PARP knock-out mice were generated by inactivation of both alleles by gene targeting. We showed that: (i) they are exquisitely sensitive to -irradiation, (ii) they died rapidly from acute radiation toxicity to the small intestine, (iii) they displayed a high genomic instability to -irradiation and MNU injection and, (iv) bone marrow cells rapidly underwent apoptosis following MNU treatment, demonstrating that PARP is a survival factor playing an essential and positive role during DNA damage recovery and survival.     

New antifungal agents that inhibit the growth of Candida Species: Dichlorinated 8-quinolinols

Five dichlorinated 8-quinolinols (2,5- 5,6-, 3,5-, 3,7-, and 4,5-dichloro-8-quinolinol) were tested against Candida albicans and C. Tropicalis in Sabouraud dextrosebroth with and without bovine serum. The 5,6-, 3,5-, and 3,7-dichloro-8-quinolinols proved to be more effective than the control, 5-fluorocytosine. In cytotoxicity tests employing baby hamster kidney (BHK) cells, all test agents proved to be more cytotoxic than the control. However, the minimum inhibitory concentration (MIC) of 3,5-dichloro-8-quinolinol to both fungi was only one tenth the cytotoxic dose,suggesting that the compound may be useful as a topical or systemic antifungal agent.This revised version was published online in October 2005 with corrections to the Cover Date.     

Selecting arthropod biological control agents against arthropod pests: Can the science be improved to decrease the risk of releasing ineffective agents?

With greater emphasis being placed on management of the risks attached to natural enemy releases for biocontrol programs and the need to justify research budgets, the efficient selection of effective natural enemies is increasingly important. Historically there has been little agreement regarding how or whether this can be accomplished. Recent studies have demonstrated that there is good correspondence between insect host-finding behavior and attack rates in well-designed laboratory studies and their performance of this behavior in the field. Success in measuring efficacy of candidate agents remains somewhat of an art due to the multitude of factors influencing efficacy, but will be improved by attention to: (1) characterization of natural enemy candidates using morphological taxonomy or genetic markers at the onset of a program, (2) climatic matching candidate agents when possible, and (3) evaluations in semi-field or field cage conditions following quarantine evaluations whenever possible before proceeding with widespread releases. The application of these principles is discussed in regard to US biocontrol programs for Bemisia tabaci (Gennadius), Lygus spp., and Aphis glycines Matsumura. Proper project planning and interdisciplinary cooperation will enhance the chances for a successful project.     

Optimization of protocols for microinjection-based delivery of cryoprotective agents into Japanese whiting Sillago japonica embryos

Microinjection has proven useful for introduction of low-permeability cryoprotective agents (CPAs) into fish eggs or embryos for cryopreservation. In this work, we examined the suitable conditions for single or combined microinjection into the perivitelline space (PS) and the yolk mass (YM) of embryos of the Japanese whiting, an alternative marine fish model for embryo cryopreservation studies. The parameters examined were injection volume, CPA type and concentration, vehicle (diluent), and suitable developmental stage. Somites and tail elongation embryos tolerated single or combined injection with 2.1 and 15.6 nl in the PS and YM, respectively, whereas earlier embryonic stages tolerated only up to 8.2 nl in the YM. The injected solutions diffused rapidly throughout the PS and YM and remained contained within each compartment unless in the case of structural damage caused by injection of larger volumes. Yamamoto solution was marginally better as a vehicle for microinjection of CPAs than fish Ringer and phosphate buffer saline whereas ¼ artificial sea water was clearly unsuitable. Ethylene glycol was well tolerated by embryos in all developmental stages whereas 1, 2-propylene glycol was suitable only for early embryonic stages. Overall, microinjection was efficient in delivering high loads of CPAs inside whiting embryos more swiftly than previously obtained for this species by immersion-based impregnation protocols. Embryos microinjected with CPAs showed a decrease in embryo nucleation temperature and an increase in chilling tolerance. CPA-microinjected embryos will provide valuable materials to optimize the remaining parameters that are critical for successful cryopreservation such as cooling and warming strategies.     

抗血管生成药物联合化疗治疗进展期非小细胞肺癌研究现状

大部分非小细胞肺癌患者需要进行系统化疗。研究表明以铂类药物为基础的标准一线系统化疗联合贝伐单抗能一定程度改善晚期非小细胞肺癌患者的预后。另有一些研究对多靶点抗血管生成的酪氨酸激酶抑制剂(如:索拉非尼,舒尼替尼,西地尼布,凡德他尼等)联合标准化疗的疗效进行了评估,为非小细胞肺癌患者提供了更多的治疗策略。本文主要对抗血管生成药物联合细胞毒性化疗药物治疗非小细胞肺癌的临床研究进行系统回顾。     

尖端赛多孢子菌引起的真菌性鼻窦炎1例并文献复习

目的 探讨尖端赛多孢子菌的实验室检测方法,了解其对5种常用抗真菌药物的体外敏感性.通过对相关文献的复习,熟悉真菌性鼻窦炎的临床表现和诊治方法.方法 将1例尖端赛多孢子菌引起的真菌性鼻窦炎患者经鼻内腔镜手术切除的团块用10％ KOH压片镜检、涂片革兰染色镜检、沙堡弱培养基培养、分子生物学方法鉴定到种.分离株应用E-test进行体外药敏试验.结果 鉴定为尖端赛多孢子菌.5种药物对其MIC范围分别为:伏立康唑0.064 μg/mL,卡泊芬净1.500 μg/mL,氟康唑16.000 μg/mL,两性霉素B＞32.000 μg/mL,5-氟胞嘧啶＞32.000 μg/mL.结论 尖端赛多孢子菌引起的真菌性鼻窦炎国内少见报道,易与肿瘤相混淆;实验室检测对正确诊断起决定性作用;行鼻内腔镜下手术治疗效果较好.了解该菌的耐药性对指导抗真菌治疗尤为关键.     

Potential CRF1R PET imaging agents: N-fluoroalkyl-8-(6-methoxy-2-methylpyridin-3-yl)-2,7-dimethyl-N-alkylpyrazolo[1,5-a][1,3,5]triazin-4-amines

A series of N-fluoroalkyl-8-(6-methoxy-2-methylpyridin-3-yl)-2,7-dimethyl-N-alkylpyrazolo[1,5-a][1,3,5]triazin-4-amines were prepared and evaluated as potential CRF₁R PET imaging agents. Optimization of their CRF₁R binding potencies and octanol-phosphate buffer phase distribution coefficients resulted in discovery of analog 7e (IC₅₀ = 6.5 nM, log D = 3.5).