Kinetics of Photobleaching of Protoporphyrin IX in the Skin of Nude Mice Exposed to Different Fluence Rates of Red Light

Abstract— The purpose of the present study was to determine the kinetics and the fluence rate dependency of the photo-bleaching of protoporphyrin IX (PpIX) in normal skin of Balb/c nude mice after systemic and topical application of 5-aminolevulinic acid (ALA). ALA was administered systemically (200 mg/kg body weight, i.p.) and topically (20% w/w ALA cream) to the mice. Fluences of up to 40 J/cm² were delivered by a dye laser (636 nm) at fluence rates of 37.5, 75, 150, 300 and 500 mW/cm². The photo-bleaching rate was constant within this range of fluence rates. This result suggests that there is no oxygen effect for PpIX photobleaching in this region for the skin of Balb/c nude mice. During light exposure the fluorescence decay followed neither first- nor second-order kinetics. The decay rate was slightly faster after systemic application than after topical application of ALA, but did not depend on the time (1–8 h) between application and analysis.     

The Effect of Fluence Rate on Tumor and Normal Tissue Responses to Photodynamic Therapy

Photodynamic therapy (PDT), carried out at low fluence rates, may enhance tumor response as well as affect treatment selectivity. We have studied the effects of fluence rate on the response of the murine radiation-induced fibrosarcoma (RIF) to PDT using Photofrin® (5 mg/kg). Tumor response was tested over a large range of fluence rates (10-200 mW/cm²) and fluences (25-378 J/ cm²). Low fluence rates were more efficient; -60 J/cm2 at 10 mW/cm² was needed to achieve the same tumor growth delay as -100 J/cm² at 150 mW/cm² and -150 J/cm2 at 200 mW/cm². Despite this increased efficiency, lower fluence rates still required longer treatment times for equivalent anti-tumor effects: 95 min for 57 J/cm² at 10 mW/cm²versus 11 min for 100 J/cm² at 150 mW/cm². Effects of fluence rate on the PDT toxicity to normal tissue were examined through the response of the murine (C311) foot to Photofrin® PDT. Treatment with conditions that produced equivalent tumor responses, i.e. 57 J/cm² at 10 mW/cm² and 100 J/cm² at 150 mW/cm², resulted in a more severe foot response at the higher fluence rate (median peak response: 0.9 at 10 mW/cm², 1.5 at 150 mW/cm²) with more time required for tissue to return to normal (8 days at 10 mW/cm², at least 30 days at 150 mW/cm²). However, when feet were treated with an equal fluence of 100 J/cm2 at various fluence rates, longer healing times accompanied the lower fluence rate treatments. Overall, this paper demonstrates that lower PDT fluence rates are associated with increased efficiency of tumor response. If this increased efficiency is accounted for by lowering treatment fluence, lower fluence rates also may result in a more favorable normal tissue response to treatment.     

Phthalocyanine 4 (Pc 4) photodynamic therapy of human OVCAR-3 tumor xenografts

Photodynamic therapy (PDT) is a cancer treatment modality utilizing a photosensitizer, light and oxygen. Photodynamic therapy with Photofrin has been approved by the U.S. Food and Drug Administration for treatment of advanced esophageal and early lung cancer. Because of certain drawbacks associated with the use of Photofrin, there is a need to identify new photosensitizers for human use. The photosensitizer Pc 4 (HOSiPc-OSi[CH3]2[CH2]3N[CH3]2) has yielded promising PDT effects in many in vitro and in vivo systems. The aim of this study was to assess the usefulness of Pc 4 as a PDT photosensitizer for a human tumor grown as a xenograft in athymic nude mice. The ovarian epithelial carcinoma (OVCAR-3) was heterotransplanted subcutaneously in athymic nude mice. Sixty mice bearing OVCAR-3 tumors (approximately 80-130 mm3) were divided into six groups of 10 animals each, three for controls and three for treatment. The Pc 4 was given by tail vein injection, and 48 h later a 1 cm area encompassing the tumor was irradiated with light from a diode laser coupled to a fiberoptic terminating in a microlens (lambda = 672 nm, 150 J/cm2, 150 mW/cm2). Tumors of control animals receiving no treatment, light alone or Pc 4 alone continued to grow. Of animals receiving 0.4 mg/kg Pc 4 and light, one (10%) had a complete response and was cured (no regrowth up to 90 days post-PDT), while all others (90%) had a partial response and were delayed in regrowth. Of animals receiving 0.6 mg/kg Pc 4 and light, eight (80%) had a complete response, and two of these were cured. Of animals receiving 1.0 mg/kg Pc 4 and light, six (60%) had a complete response, and two of these were cured. In additional experiments, tumors from animals treated with Pc 4 (1 mg/kg) and light were removed 15, 30, 60 and 180 min post-PDT, and from these tumors DNA and protein were extracted. Agarose gel electrophoresis revealed the presence of apoptotic DNA fragmentation as early as 15 min post-PDT. Western blotting showed the cleavage of the 116 kDa native poly(ADP-ribose) polymerase (PARP) into fragments of approximately 90 kDa, another indication of apoptosis, and the presence of p21/WAF1/CIP1 (p21) in all PDT-treated tumors. These changes did not occur in control tumors. Pc 4 appears to be an effective photosensitizer for PDT of human tumors grown as xenografts in nude mice. Early apoptosis, as revealed by PARP cleavage, DNA fragmentation and p21 overexpression, may be responsible for the excellent Pc 4-PDT response. Clinical trials of Pc 4-PDT are warranted.     

Elemental Selenium Generated by the Photobleaching of Seleno-Merocyanine Photosensitizers Forms Conjugates with Serum Macro-Molecules That are Toxic to Tumor Cells

Abstract

Elemental selenium generated by the photobleaching of selenomerocyanine dyes forms conjugates with serum albumin and serum lipoproteins that are toxic to leukemia and selected solid tumor cells but well tolerated by normal CD34-positive hematopoietic stem and progenitor cells. Serum albumin and lipoproteins act as Trojan horses that deliver the cytotoxic entity (elemental selenium) to tumor cells as part of a physiological process. They exploit the fact that many tumors have an increased demand for albumin and/or low-density lipoprotein. Se(0)-protein conjugates are more toxic than selenium dioxide, sodium selenite, selenomethionine, or selenocystine. They are only minimally affected by a drug resistance mechanism, and they potentiate the cytotoxic effect of ionizing radiation and several standard chemotherapeutic agents. The cytotoxic mechanism of Se(0)-protein conjugates is not yet fully understood. Currently available data are consistent with the notion that Se(0)-protein conjugates act as air oxidation catalysts that cause a rapid depletion of intracellular glutathione and induce apoptosis. Drugs modeled after our Se(0)-protein conjugates may prove useful for the local and/or systemic therapy of cancer.     

A Comparison of Dose Metrics to Predict Local Tumor Control for Photofrin‐mediated Photodynamic Therapy

This preclinical study examines light fluence, photodynamic therapy (PDT) dose and “apparent reacted singlet oxygen,” [¹O₂]_rx, to predict local control rate (LCR) for Photofrin‐mediated PDT of radiation‐induced fibrosarcoma (RIF) tumors. Mice bearing RIF tumors were treated with in‐air fluences (50–250 J cm^?2) and in‐air fluence rates (50–150 mW cm^?2) at Photofrin dosages of 5 and 15 mg kg^?1 and a drug‐light interval of 24 h using a 630‐nm, 1‐cm‐diameter collimated laser. A macroscopic model was used to calculate [¹O₂]_rx and PDT dose based on in vivo explicit dosimetry of the drug concentration, light fluence and tissue optical properties. PDT dose and [¹O₂]_rx were defined as a temporal integral of drug concentration and fluence rate, and singlet oxygen concentration consumed divided by the singlet oxygen lifetime, respectively. LCR was stratified for different dose metrics for 74 mice (66 + 8 control). Complete tumor control at 14 days was observed for [¹O₂]_rx ≥ 1.1 mm or PDT dose ≥1200 μm J cm^?2 but cannot be predicted with fluence alone. LCR increases with increasing [¹O₂]_rx and PDT dose but is not well correlated with fluence. Comparing dosimetric quantities, [¹O₂]_rx outperformed both PDT dose and fluence in predicting tumor response and correlating with LCR.     

Quantitative analysis of Pc 4 localization in mouse lymphoma (LY-R) cells via double-label confocal fluorescence microscopy

Photodynamic therapy (PDT) is a novel cancer therapy that uses light-activated drugs (photosensitizers) to destroy tumor tissue. Reactive oxygen species produced during PDT are thought to cause the destruction of tumor tissue. However, the precise mechanism of PDT is not completely understood. To provide insight into the in vitro mechanisms of PDT, we studied the subcellular localization of the photosensitizer HOSiPcOSi(CH3)2-(CH2)3N(CH3)2 (Pc 4) in mouse lymphoma (LY-R) cells using double-label confocal fluorescence microscopy. This technique allowed us to observe the relative distributions of Pc 4 and an organelle-specific dye within the same cell via two, spectrally distinct, fluorescence images. To quantify the localization of Pc 4 within different organelles, linear correlation coefficients from the fluorescence data of Pc 4 and the organelle-specific dyes were calculated. Using this measurement, the subcellular spatial distributions of Pc 4 could be successfully monitored over an 18 h period. At early times (0-1 h) after introduction of Pc 4 to LY-R cells, the dye was found in the mitochondria, lysosomes and Golgi apparatus, as well as other cytoplasmic membranes, but not in the plasma membrane or the nucleus. Over the next 2 h, there was some loss of Pc 4 from the lysosomes as shown by the correlation coefficients. After an additional incubation period of 2 h Pc 4 slowly increased its accumulation in the lysosomes. The highest correlation coefficient (0.65) was for Pc 4 and BODIPY-FL C5 ceramide, which targets the Golgi apparatus, and also binds to other cytoplasmic membranes. The correlation coefficient was also high (0.60) for Pc 4 and a mitochondria-targeting dye (Mitotracker Green FM). Both of these correlation coefficients were higher than that for Pc 4 with the lysosome-targeting dye (Lysotracker Green DND-26). The results suggest that Pc 4 binds preferentially and strongly to mitochondria and Golgi complexes.     

Inhibition of various steps in the replication cycle of vesicular stomatitis virus contributes to its photoinactivation by AlPcS4 or Pc4 and red light

Vesicular stomatitis virus (VSV) was used as a model virus to study the processes involved in photoinactivation by aluminum phthalocyanine tetrasulfonate (AlPcS4) or silicon phthalocyanine HOSiPcOSi(CH3)2(CH2)3N(CH3)2 (Pc4) and red light. Previously a very rapid decrease in the intracellular viral RNA synthesis after photodynamic treatment was observed. This decrease was correlated to different steps in the replication cycle. Binding of VSV to host cells and internalization were only slightly impaired and could be visualized by electron microscopy. The capability of the virus to fuse with membranes in an acidic endosomal environment was studied using both pyrene-labeled liposomes and a hemolysis assay as a model. These tests indicate a rapid decrease of fusion capacity after AlPcS4 treatment, which correlated with the decrease in RNA synthesis. For Pc4 treatment no such correlation was found. The fusion process is the first step in the replication cycle, affected by AlPcS4 treatment, but also in vitro RNA polymerase activity was previously shown to be inhibited. Inactivation of VSV by Pc4 treatment is apparently caused by damage to a variety of viral components. Photodynamic treatment of virus suspensions with both sensitizers causes formation of 8-oxo-7,8-dihydroguanosine in viral RNA as measured by HPLC with electrochemical detection. This damage might be partly responsible for inhibition of the in vitro viral RNA polymerase activity by photodynamic treatment.     

Synthesis of 2-Heterocyclomethyl-5-diphenylmethylenecyclopentanone Hydrochlorides and their Inhibitory Effect on Tumor Cell Growth

A large number of natural diterpenes and sesquiterpenes have remarkable bioactivity. The presence of α-methylenec?yclopentanone in diterpenes and α-methylene-γ-lactones in sesquiterpenes plays a significant role to mediate anti-tumour activities by directly interacting with sulfhydryl enzymes. However, it has been recognized that the therapeutic value of these natural compounds was limited due to their indiscriminating toxicity. To increase the selectivity and cytotoxicity to tumor cells, w…     

Rapid Tyrosine Phosphorylation of HS1 in the Response of Mouse Lymphoma L5178Y-R Cells to Photodynamic Treatment Sensitized by the Phthalocyanine Pc 4

Abstract— The ability of photodynamic treatment (PDT) with the phthalocyanine Pc 4 to activate cellular signal transduction pathways in murine lymphoma L5178Y-R cells has been assessed by observing increases in protein tyrosine phosphorylation at early times post-PDT. Western blot analysis with an anti-phosphotyrosine antibody revealed a dramatic increase in phosphorylation of two major protein bands of Mr -80000 and -55000 in response to PDT. The increase was PDT dose-dependent, occurred as early as 20 s after initiation of light exposure of Pc 4-pre-loaded cells and was amplified by the presence of the protein tyrosine phosphatase inhibitor, sodium ortho-vanadate (NaV0₄). By immunoprecipitation, one of the Mr –80000 phosphorylated proteins has been identified as HS1, a substrate of nonreceptor-type protein tyrosine kinases. Although vanadate greatly enhanced the level and extent of PDT-induced phosphorylation, it had no influence on overall photocytotoxicity or on the rate of apoptotic DNA fragmentation. Genistein, an inhibitor of protein tyrosine kinases, diminished tyrosine phosphorylation of the Mr –80000 and other proteins and dramatically potentiated cell killing induced by PDT but did not significantly affect PDT-induced apoptosis. The results suggest that PDT rapidly activates a membrane-associated src family kinase(s) in L5178Y-R cells, one substrate of which is HS1, and that protein tyrosine phosphorylation is part of a stress response, protecting a portion of the cells from the lethal effects of PDT but not altering the mechanism by which they die.     

钒酸钇双折射晶体生长及缺陷观察

采用在较低氧分压的保护气氛中用提拉法(Cz法)生长YVO4晶体, 采用自行设计的气压计, 精密调节炉内的氧、氮比例, 有效防止了晶体生长中的过度缺氧, 生长出Ф33 mm×31 mm(等径)YVO4晶体. 设计了生长YVO4晶体最佳工艺条件转速5～10 r*min-1, 拉速2～6 mm*h-1, 生长周期24 h, 液面上8 mm温度梯度2.875 ℃*mm-1. 用偏光显微镜对YVO4晶体的裂纹、散射颗粒、包裹物、偏心生长等缺陷进行观察, 认为它们的成因主要是生长速率过快, 生长环境中湿度大及晶体中存在分解和挥发性物质等.     

Semi-Synthesis of Small Molecules of Aminocarbazoles: Tumor Growth Inhibition and Potential Impact on p53

The tumor suppressor p53 is inactivated by mutation in approximately 50% of human cancers. Small molecules that bind and stabilize those mutants may represent effective anticancer drugs. Herein, we report the tumor cell growth inhibitory activity of carbazole alkaloids and amino derivatives, as well as their potential activation of p53. Twelve aminocarbazole alkaloids were semi-synthesized from heptaphylline (1), 7-methoxy heptaphylline (2), and 7-methoxymukonal (3), isolated from Clausena harmandiana, using a reductive amination protocol. Naturally-occurring carbazoles 1–3 and their amino derivatives were evaluated for their potential effect on wild-type and mutant p53 activity using a yeast screening assay and on human tumor cell lines. Naturally-occurring carbazoles 1–3 showed the most potent growth inhibitory effects on wild-type p53-expressing cells, being heptaphylline (1) the most promising in all the investigated cell lines. However, compound 1 also showed growth inhibition against non-tumor cells. Conversely, semi-synthetic aminocarbazole 1d showed an interesting growth inhibitory activity in tumor cells expressing both wild-type and mutant p53, exhibiting low growth inhibition on non-tumor cells. The yeast assay showed a potential reactivation of mutant p53 by heptaphylline derivatives, including compound 1d. The results obtained indicate that carbazole alkaloids may represent a promising starting point to search for new mutp53-reactivating agents with promising applications in cancer therapy.     

Nd:GdVO4晶体的生长与光谱性能

采用Czochralski法生长了均匀透明的Nd:GdVO4晶体,测量了其室温偏振吸收谱,并与0.2mol·L^-1的NdCl3溶液的室温吸收谱进行了比较。根据Judd-Ofelt理论,拟合出晶体场唯象强度参数：Ω2=12.629×10^-20cm^2,Ω4=4.828×10^-20cm^2,Ω6=8.425×10^-20cm^2.计算了各能级的辐射跃迁几率AJ,J',振子强度PJ,J;,辐射寿命τ,荧光分支比βJ'等光学参量。并用LD激光器泵浦Nd:GVO4晶体,当量大泵浦功率为26W时,得到输出功率为14.3W波长为1.06μm的基频光,光-光转换效率为55%,斜效率为63%。     

Nucleation and Growth of CeF3 and NaCeF4 Nanocrystals

Studying growth : The diffusion‐controlled kinetic (DCK) model and the surface chemical thermodynamics (SCT) model have been successfully applied to interpret the nucleation and growth mechanism of CeF₃ (see TEM images) and NaCeF₄ nanocrystals, and may generally give light to the size‐control and morphology prediction of rare‐earth fluorides at the nanoscale level.

     

八面体纳米钼酸钡的原位生长及形成机理

利用RD496-2000型微量热计获取的微纳钼酸钡原位生长的热动力学信息解释了微纳钼酸钡制备反应的热动力学机理; 微纳钼酸钡制备反应均经历放热-吸热-放热的表观过程, 表观速率常数k和表观反应级数n变化趋势相同, 但数值和变化率存在差异; 通过对比分析出微乳液体系与水体系反应的量热差异与反胶束的碰撞、传递、融合、分离和重组等复杂过程密切相关.     

Growth and Crystal Structure of YbAl3（BO3）4

1 INTRODUCTION With the development of high power InGaAs laser diode (LD) as pump sources, the Yb3+-doped laser crystals have received much attention. Due to the simple energy level structure of the Yb3+ ions, the Yb3+-doped laser crystals have no concentration quenching, and have high quantum efficiency and low quantum defects even at high doping concentration of the Yb3+ ions[1, 2]. Therefore, the Yb3+-doped crystals are good candidates of media in the LD-pumped solid-state laser,…     

微量量热法研究螺旋藻活性物质对癌细胞和正常细胞的作用

螺旋藻对人体生理的作用已引起人们的重视,特别是在增强免疫力和抗癌方面。本文用标准型(MS 80)Calvet微量量热计测定了正常生长状况下人宫颈癌传代细胞(Hela)、人乳腺癌细胞(Bcap-37)和正常人胚肺二倍体细胞(2BS)的增殖热谱图;螺旋藻活性物质(Sp,P)对上述癌细胞和正常细胞作用的热谱图。同时,测定了量热实验前后的细胞数、Sp.S对细胞生长的抑制率和细胞死亡率。     

气氛对MnCr2O4尖晶石纳米线生长的影响

建立了一种在氮气和氢气的还原性混合气氛和1100 ℃条件下加热商业不锈钢箔(304)制备MnCr2O4尖晶石纳米线的简单方法, 并研究了不同气氛对纳米线生长的影响. 研究发现, 混合气体中氢气含量的变化会影响纳米线的形貌和产率; 而氧化性气氛(如空气)下则得不到纳米线. 在还原性气氛下, Mn和Cr原子可以和反应室内残留的痕量氧反应生成MnCr2O4尖晶石, 而Fe和Ni原子不能被氧化, 但是Fe和Ni可以起到催化纳米线生长的作用, 纳米线的生长机理属于自催化性的气-液-固(VLS)机制.     

A Macrocyclic Ruthenium(III) Complex Inhibits Angiogenesis with Down‐Regulation of Vascular Endothelial Growth Factor Receptor‐2 and Suppresses Tumor Growth In Vivo

A macrocyclic ruthenium(III) complex [Ru^III(N₂O₂)Cl₂]Cl ( Ru‐1 ) is reported as an inhibitor of angiogenesis and an anti‐tumor compound. The complex is relatively non‐cytotoxic towards endothelial and cancer cell lines in vitro, but specifically inhibited the processes of angiogenic endothelial cell tube formation and cancer cell invasion. Moreover, compared with known anti‐cancer ruthenium complexes, Ru‐1 is distinct in that it suppressed the expression of vascular endothelial growth factor receptor‐2 (VEGFR2), and the associated downstream signaling that is crucial to tumor angiogenesis. In addition, in vivo studies showed that Ru‐1 inhibited angiogenesis in a zebrafish model and suppressed tumor growth in nude mice bearing cancer xenografts.     

Growth mechanism of penniform BaWO4 nanostructures in catanionic reverse micelles involving polymers

The formation of penniform BaWO4 nanostructures made of nanowires or nanobelts under the direction of a block copolymer in catanionic reverse micelles has been studied in detail. On the basis of the experimental results obtained from the BaWO4 crystallization in aqueous polymer solutions and careful transmission electron microscopy (TEM) observations of BaWO4 nanostructures formed in reverse micelles containing polymers, a detailed two-stage growth mechanism has been proposed for the formation of the penniform nanostructures in reverse micelles, which involves the polymer-controlled shaft formation (Stage 1) and the mixed surfactants-controlled barb growth (Stage 2). During Stage 1, poly(ethylene glycol)-block-poly(methacrylic acid) (PEG-b-PMAA) induced the formation of c-axis-oriented shuttle-like nanocrystals and the subsequent oriented attachment of these shuttle-like nanocrystals resulted in the formation of [100]-oriented shafts with many parallel [001]-oriented pricks. During Stage 2, [001]-oriented nanowires or nanobelts grew gradually from the pricks into barbs, leading to the formation of well-defined penniform BaWO4 nanostructures with the barb morphology essentially determined by the mixing ratio r of the anionic to cationic surfactants (i.e., nanowires were formed at r=1 while nanobelts were formed at r deviating from 1). The current understanding of the growth mechanism of penniform BaWO4 nanostructures in catanionic reverse micelles involving polymers may be potentially applied for designing a new synthesis system for the controlled synthesis of other hierarchical 1D nanostructures with desired architectures.