Intracellular localization of the antitumour drug adriamycin in living cultured cells: A confocal microscopy study

The intracellular distribution of the anthracyclinic antibiotic adriamycin in living cultured cells has been investigated by confocal microscopy. In human melanoma cells (M14), adriamycin was localized inside the nuclei. When adriamycin-treated M14 cells were allowed to recover in drug-free medium, a complete efflux of the drug from the nucleus was revealed. In recovered cells, a weakly fluorescent signal was observed in the perinuclear region. When M14 cells were recovered in a medium containing colcemid, a microtubule depolymerizing agent, the drug transport from the nucleus to the cell periphery appeared to be inhibited, suggesting that the microtubule network is strongly involved in drug transport mechanisms. In multidrug-resistant (MDR) cells the intracellular location of adriamycin was shown to be noticeably different from that of the parental wild-type cells. In particular, in resistant human breast carcinoma cells (MCF-7), adriamycin appeared to be exclusively located within the cytoplasm whereas the nuclei were shown to be completely negative. When adriamycin treatment was performed in association with MDR revertants, such as Lonidamine (inhibitor of the energy metabolism) or verapamil (inhibitor of the P-glycoprotein efflux pump), a marked enhancement of the cytoplasmic signal was observed in resistant cells. Under these conditions, adriamycin appeared concentrated in the perinuclear region, but the nuclei were still negative. Confocal microscopy proved to be a very useful method for the study of the intracellular transport of fluorescent substances, such as anthracyclinic antibiotics, and for the investigation of the multidrug resistance phenomenon in tumour cells.     

Prospects and Challenges of the Study of Anti-Glycan Antibodies and Microbiota for the Monitoring of Gastrointestinal Cancer

Over the past decades, a large amount of data has been accumulated in various subfields of glycobiology. However, much clinically relevant data and many tools are still not widely used in medicine. Synthetic glycoconjugates with the known structure of glycans are an accurate tool for the study of glycan-binding proteins. We used polyacrylamide glycoconjugates (PGs) including PGs with tumour-associated glycans (TAGs) in immunoassays to assess the prognostic potential of the serum level of anti-glycan antibodies (AG Abs) in gastrointestinal cancer patients and found an association of AG Abs with survival. The specificity of affinity-isolated AG Abs was investigated using synthetic and natural glycoconjugates. AG Abs showed mainly a low specificity to tumour-associated and tumour-derived mucins; therefore, the protective role of the examined circulating AG Abs against cancer remains a challenge. In this review, our findings are analysed and discussed in the context of the contribution of bacteria to the AG Abs stimulus and cancer progression. Examples of the influence of pathogenic bacteria colonising tumours on cancer progression and patient survival through mechanisms of interaction with tumours and dysregulated immune response are considered. The possibilities and problems of the integrative study of AG Abs and the microbiome using high-performance technologies are discussed.     

Stimuli-Responsive Polymeric Nanomaterials for the Delivery of Immunotherapy Moieties: Antigens,Adjuvants and Agonists

Immunotherapy has been investigated for decades, and it has provided promising results in preclinical studies. The most important issue that hinders researchers from advancing to clinical studies is the delivery system for immunotherapy agents, such as antigens, adjuvants and agonists, and the activation of these agents at the tumour site. Polymers are among the most versatile materials for a variety of treatments and diagnostics, and some polymers are reactive to either endogenous or exogenous stimuli. Utilizing this advantage, researchers have been developing novel and effective polymeric nanomaterials that can deliver immunotherapeutic moieties. In this review, we summarized recent works on stimuli-responsive polymeric nanomaterials that deliver antigens, adjuvants and agonists to tumours for immunotherapy purposes.     

The journal Communications in Numerical Methods in Engineering with Biomedical Applications becomes the International Journal for Numerical Methods in Biomedical Engineering (IJNMBE) from 1st January 2010

Communications in Numerical Methods in Engineering, founded by Roland W. Lewis in 1985, will change its title to the ‘International Journal for Numerical Methods in Biomedical Engineering’ and have a revised Aims and Scope. Copyright © 2009 John Wiley & Sons, Ltd.     

Structure-activity studies of human tumour necrosis factors

The mechanism by which tumour necrosis factors (TNF and lymphotoxin,also called TNFß and TNFß respectively) exerttheir cytotoxic activity on many malignant cells, remains largelyunknown. Furthermore, the broad array of differentiation (geneinduction) and mitogenic activities towards many primary cellsis still a subject of intensive investigation. TNF is an importantmediator in inflammation, immune responses and infection-relatedphenomena and these activities contribute to the severe toxidtyseen when TNF is used as an anticancer agent. The first stepin the mechanism of action is the specific binding of the ligandto its receptors and dissection of the molecular mechanism involvedin this interaction is the subject of this review. The reasonsfor the interest in this aspect are obvious: first, the developmentof strong antagonistic TNF analogues can be useful in dampeningthe potentially lethal or debilitating effects of an overproductionof the cytokine (as in septic shock or rheumatoid arthritis).Secondly, since two distinct TNF receptors exist, constructionof TNF muteins that distinguish between both types may leadto derivatives of this plekrtropic agent with a more restrictedbiological activity pattern. Ideally, one would like to developa TNF mutant that has retained its cytotoxic action on tumourcells without inducing the deleterious systemic toxteity. Suchan optimized TNF molecule could become a potent anticancer agent     

Analysis of boron-10 in soft tissue by dynamic secondary ion mass spectrometry

We report here a preliminary study in which dynamic secondary ion mass spectrometry (SIMS) has provided images of boron‐10 (¹⁰B) in biological tissue as used in research into boron neutron capture therapy. Cultured tumour cells incubated in media containing known concentrations of a ¹⁰B‐containing compound, p‐boronophenylalanine (BPA), and intracranial tumour tissue from animals previously injected with BPA were analysed by an in‐house constructed SIMS. Investigations were conducted in positive secondary ion detection mode using a 25‐keV, 5‐nA gallium primary ion source. For calibration purposes, tissue standards were also analysed and their boron‐to‐carbon signal ratios correlated to bulk boron concentrations measured by inductively coupled plasma atomic emission spectroscopy (ICP‐AES). Ion maps of ¹⁰B, ¹²C, ²³Na and ³⁹K showing gross tissue and cell features were acquired. SIMS and ICP‐AES standard measurements were in good agreement. Tissue regions with high or low ¹⁰B concentrations were identified along with ¹⁰B hotspots in normal brain areas. Cultured cells revealed the intracellular localization of ¹⁰B. SIMS is capable of producing images showing the distribution of ¹⁰B at p.p.m. levels in cells and in normal and tumour‐bearing brain tissue.     

大鼠急性重症梗阻性黄疸减黄过程中TNF-α和IL-2的变化

目的探讨大鼠急性重症梗阻性黄疸减黄过程中血清TNF-α和IL-2及肝损伤的相关性,术前减黄对于免疫功能恢复的意义。方法以结扎胆总管（LCD）建立阻黄模型（LCD组,n=10）,在建立阻黄模型1周后松解胆总管（ID组,n=60）,正常组不进行任何处理（n=10）。采用放射免疫法测定正常组、LCD组术后1周及ID组不同时间点TNF-α、IL-2的含量,同时自动生化分析仪检测ALT和胆红素含量。结果 LCD组血清TNF-α为（1 320.58±82.15）μg.L-1,IL-2为（45.12±12.65）μg.L-1,与正常组相比差异均有统计学意义（P〈0.01）;引流1周时ALT和胆红素分别为（7.25±1.16）U.L-1和（51.03±7.52）μmol.L-1,与LCD组比较差异有统计学意义（P〈0.01）,与正常组比较差异无统计学意义（P〉0.01）;TNF-α在引流2周时已恢复正常,IL-2在引流4周时已恢复正常,与正常组比较差异均无统计学意义（P〉0.01）;LCD组和引流组不同时间点的TNF-α与ALT呈正相关（r=0.762,P〈0.01）。结论 TNF-α和IL-2参与了肝损伤,术前减黄治疗对于急性重症梗阻性黄疸免疫功能的恢复具有重要意义。