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Feng-Yun J Huang,1 Te-Wei Lee,2 Chih-Hsien Chang,2 Liang-Cheng Chen,2 Wei-Hsin Hsu,2 Chien-Wen Chang,1 Jem-Mau Lo1 1Department of Biomedical Engineering and Environmental Sciences, National Tsing Hua University, Hsinchu, Taiwan; 2Institute of Nuclear Energy Research, Longtan, Taiwan Purpose: In this study, the 188Re-labeled PEGylated nanoliposome (188Re-liposome) was prepared and evaluated as a therapeutic agent for glioma.Materials and methods: The reporter cell line, F98luc was prepared via Lentivector expression kit system and used to set up the orthotopic glioma-bearing rat model for non-invasive bioluminescent imaging. The maximum tolerated dose applicable in Fischer344 rats was explored via body weight monitoring of the rats after single intravenous injection of 188Re-liposome with varying dosages before the treatment study. The OLINDA/EXM 1.1 software was utilized for estimating the radiation dosimetry. To assess the therapeutic efficacy, tumor-bearing rats were intravenously administered 188Re-liposome or normal saline followed by monitoring of the tumor growth and animal survival time. In addition, the histopathological examinations of tumors were conducted on the 188Re-liposome-treated rats.Results: By using bioluminescent imaging, the well-established reporter cell line (F98luc) showed a high relationship between cell number and its bioluminescent intensity (R2=0.99) in vitro; furthermore, it could also provide clear tumor imaging for monitoring tumor growth in vivo. The maximum tolerated dose of 188Re-liposome in Fischer344 rats was estimated to be 333 MBq. According to the dosimetry results, higher equivalent doses were observed in spleen and kidneys while very less were in normal brain, red marrow, and thyroid. For therapeutic efficacy study, the progression of tumor growth in terms of tumor volume and/or tumor weight was significantly slower for the 188Re-liposome-treated group than the control group (P

Objective: A recombinant annexin A5 with the N-terminal extension of six histidine residues was labeled with (99m)Tc(I)-tricarbonyl ion to produce the (99m)Tc-labeled annexin A5, referred to (99m)Tc(I)-his(6)-annexin A5. We have explored the agent as an effective imaging probe for in vivo detecting the apoptosis of internal tissue subjected with high radiation doses in a γ-irradiated mouse model. Methods: [(99m)Tc(CO)(3)(OH(2))(3)](+) was prepared and taken to directly label his(6)-annexin A5. The radiochemical purity of (99m)Tc(I)-his(6)-annexin A5 after size-exclusion separation was measured by HPLC. The binding affinity of (99m)Tc(I)-his(6)-annexin A5 to apoptotic cells was assessed using 20 Gy-irradiated Jurkat T cells. The effectiveness of (99m)Tc(I)-his(6)-annexin A5 as an imaging probe to detect the internal tissue apoptosis was assessed by biodistribution study and nanoSPECT/CT using the animal model of C57BL/6J mice conducted with 10 Gy γ irradiation. Results: The radiochemical purity of (99m)Tc(I)-his(6)-annexin A5 could attain ≥95%. The binding affinity of (99m)Tc(I)-his(6)-annexin A5 to the 20 Gy-irradiated Jurkat cells was found to be ca. 20-fold higher than that to the sham-irradiated cells. In the animal imaging study, the splenic uptake of (99m)Tc(I)-his(6)-annexin A5 for the 10 Gy-irradiated mice showed from ca. 3-fold to 5-fold higher than those of the sham-irradiated mice from 45 to 165 min postinjection. The corresponding intestinal uptake showed from ca. 2-fold to 3-fold higher during the same period of time postinjection. The biodistribution study demonstrated the organ uptakes comparable with the imaging results. The apoptotic extents of the spleen and the intestine from the SPECT/CT imaging were correlated with an immunohistochemical staining assay for caspase 3 active form fragment. Conclusion: This work is the first study to demonstrate that (99m)Tc(I)-his(6)-annexin A5 is a potential clinical imaging agent for detecting radiation-induced tissue apoptosis in an animal model. [ABSTRACT FROM AUTHOR]

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection continues to pose threats to public health. The clinical manifestations of lung pathology in COVID-19 patients include sustained inflammation and pulmonary fibrosis. The macrocyclic diterpenoid ovatodiolide (OVA) has been reported to have anti-inflammatory, anti-cancer, anti-allergic, and analgesic activities. Here, we investigated the pharmacological mechanism of OVA in suppressing SARS-CoV-2 infection and pulmonary fibrosis in vitro and in vivo. Our results revealed that OVA was an effective SARS-CoV-2 3CLpro inhibitor and showed remarkable inhibitory activity against SARS-CoV-2 infection. On the other hand, OVA ameliorated pulmonary fibrosis in bleomycin (BLM)-induced mice, reducing inflammatory cell infiltration and collagen deposition in the lung. OVA decreased the levels of pulmonary hydroxyproline and myeloperoxidase, as well as lung and serum TNF-ɑ, IL-1β, IL-6, and TGF-β in BLM-induced pulmonary fibrotic mice. Meanwhile, OVA reduced the migration and fibroblast-to-myofibroblast conversion of TGF-β1-induced fibrotic human lung fibroblasts. Consistently, OVA downregulated TGF-β/TβRs signaling. In computational analysis, OVA resembles the chemical structures of the kinase inhibitors TβRI and TβRII and was shown to interact with the key pharmacophores and putative ATP-binding domains of TβRI and TβRII, showing the potential of OVA as an inhibitor of TβRI and TβRII kinase. In conclusion, the dual function of OVA highlights its potential for not only fighting SARS-CoV-2 infection but also managing injury-induced pulmonary fibrosis., Competing Interests: Conflict of interest statement The authors declare no conflict of interest., (Copyright © 2023. Published by Elsevier Masson SAS.)

A major challenge to end the pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is to develop a broadly protective vaccine that elicits long-term immunity. As the key immunogen, the viral surface spike (S) protein is frequently mutated, and conserved epitopes are shielded by glycans. Here, we revealed that S protein glycosylation has site-differential effects on viral infectivity. We found that S protein generated by lung epithelial cells has glycoforms associated with increased infectivity. Compared to the fully glycosylated S protein, immunization of S protein with N-glycans trimmed to the mono-GlcNAc-decorated state (S MG ) elicited stronger immune responses and better protection for human angiotensin-converting enzyme 2 (hACE2) transgenic mice against variants of concern (VOCs). In addition, a broadly neutralizing monoclonal antibody was identified from S MG -immunized mice that could neutralize wild-type SARS-CoV-2 and VOCs with subpicomolar potency. Together, these results demonstrate that removal of glycan shields to better expose the conserved sequences has the potential to be an effective and simple approach for developing a broadly protective SARS-CoV-2 vaccine.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection causes severe "flu-like" symptoms that can progress to acute respiratory distress syndrome (ARDS), pneumonia, renal failure, and death. From the therapeutic perspective, 3-chymotrypsin-like protein (3CLpro) is a plausible target for direct-acting antiviral agents because of its indispensable role in viral replication. The flavonoid ugonin J (UJ) has been reported to have antioxidative and anti-inflammatory activities. However, the potential of UJ as an antiviral agent remains unexplored. In this study, we investigated the therapeutic activity of UJ against SARS-CoV-2 infection. Importantly, UJ has a distinct inhibitory activity against SARS-CoV-2 3CLpro, compared to luteolin, kaempferol, and isokaempferide. Specifically, UJ blocks the active site of SARS-CoV-2 3CLpro by forming hydrogen bonding and van der Waals interactions with H163, M165 and E166, G143 and C145, Q189, and P168 in subsites S1, S1', S2, and S4, respectively. In addition, UJ forms strong, stable interactions with core pharmacophore anchors of SARS-CoV-2 3CLpro in a computational model. UJ shows consistent anti-inflammatory activity in inflamed human alveolar basal epithelial A549 cells. Furthermore, UJ has a 50% cytotoxic concentration (CC 50 ) and a 50% effective concentration (EC 50) values of about 783 and 2.38 µM, respectively, with a selectivity index (SI) value of 329, in SARS-CoV-2-infected Vero E6 cells. Taken together, UJ is a direct-acting antiviral that obstructs the activity of a fundamental protease of SARS-CoV-2, offering the therapeutic potential for SARS-CoV-2 infection., Competing Interests: Author YW was employed by the company Arjil Biotech Holding Company Limited. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Chiou, Lu, Hsu, Chang, Chin, Liu, Lo, Wu, Yang and Huang.)

Balloon angioplasty-induced neointimal hyperplasia remains a clinical problem that must be resolved. The bioactivities of the Crossostephium chinense extract (CCE) have demonstrated potential in preventing the progression of restenosis. The present study evaluated whether CCE can suppress balloon angioplasty-induced neointima formation and elucidated its possible pharmacological mechanisms. A rat model of carotid arterial balloon angioplasty was established to evaluate the inhibitory effect of CCEs on neointimal hyperplasia. Two cell lines, A10 vascular smooth muscle cells (VSMCs) and RAW264.7 macrophages, were used to investigate the potential regulatory activities and pharmacological mechanisms of CCEs in cell proliferation and migration and in inflammation. Our in vitro results indicated that CCE3, the ethanolic extract of C. chinense , exerted the strongest growth inhibitory and antimigratory effects on VSMCs. CCE3 blocked the activation of focal adhesion kinase, platelet-derived growth factor receptor- β (PDGFRB), and its downstream molecules (AKT and mTOR) and reduced the expression of matrix metalloproteinase-2. In addition, our findings revealed that CCE3 significantly increased the expression of miRNA-132, an inhibitory regulator of inflammation and restenosis, and suppressed the expression of inflammation-related molecules (inducible nitric oxide synthase, cyclooxygenase-2, interleukin- (IL-) 1 β , and IL-6). Our in vivo study results indicated that balloon injury-induced neointimal hyperplasia was inhibited by CCE3. CCE3 could reduce neointima formation in balloon-injured arteries, and this effect may be partially attributed to the CCE3-induced suppression of PDGFRB-mediated downstream pathways and inflammation-related molecules., Competing Interests: The authors declare that they have no conflicts of interest., (Copyright © 2021 Chun-Hsu Pan et al.)

The influenza virus hemagglutinin (HA) and coronavirus spike (S) protein mediate virus entry. HA and S proteins are heavily glycosylated, making them potential targets for carbohydrate binding agents such as lectins. Here, we show that the lectin FRIL, isolated from hyacinth beans (Lablab purpureus), has anti-influenza and anti-SARS-CoV-2 activity. FRIL can neutralize 11 representative human and avian influenza strains at low nanomolar concentrations, and intranasal administration of FRIL is protective against lethal H1N1 infection in mice. FRIL binds preferentially to complex-type N-glycans and neutralizes viruses that possess complex-type N-glycans on their envelopes. As a homotetramer, FRIL is capable of aggregating influenza particles through multivalent binding and trapping influenza virions in cytoplasmic late endosomes, preventing their nuclear entry. Remarkably, FRIL also effectively neutralizes SARS-CoV-2, preventing viral protein production and cytopathic effect in host cells. These findings suggest a potential application of FRIL for the prevention and/or treatment of influenza and COVID-19., Competing Interests: Declaration of Interests A patent application has been submitted by Academia Sinica based on the results shown in this study, with C.M., J.-T.J., and Y.-M.L. as inventors., (Copyright © 2020 The Authors. Published by Elsevier Inc. All rights reserved.)

Antrodia camphorata, a well-known and highly valued edible medicinal mushroom with intriguing activities like liver protection, has been traditionally used for the treatment of alcoholic liver disease. A. camphorata shows highly medicinal and commercial values with the demand far exceeds the available supply. Thus, the petri-dish cultured A. camphorata (PDCA) is expected to develope as a substitute. In this paper, nineteen triterpenes were isolated from PDCA, and thirteen of them were the unique anthroic acids in A. camphorata, including the main content antcin K, which suggested that PDCA produced a large array of the same anthroic acids as the wild one. Furthermore, no obvious acute toxicity was found suggesting the edible safety of PDCA. In mice alcohol-induced liver injury model, triglyceride (TG), aspartate aminotransferase (AST), alanine aminotransferase (ALT), and malondialdehyde (MDA) had been reduced by the PDCA powder as well as the main content antcin K, which indicated that the PDCA could protect alcoholic liver injury in mice model and antcin K could be the effective component responsible for the hepatoprotective activities of PDCA against alcoholic liver diseases., (Copyright © 2019 China Pharmaceutical University. Published by Elsevier B.V. All rights reserved.)

Unimolecular dissociation of 1,3,5-trioxane was investigated experimentally and theoretically over a wide range of conditions. Experiments were performed behind reflected shock waves over the temperature range of 775-1082 K and pressures near 900 Torr using a high-repetition rate time of flight mass spectrometer (TOF-MS) coupled to a shock tube (ST). Reaction products were identified directly, and it was found that formaldehyde is the sole product of 1,3,5-trioxane dissociation. Reaction rate coefficients were extracted by the best fit to the experimentally measured concentration-time histories. Additionally, high-level quantum chemical and RRKM calculations were employed to study the falloff behavior of 1,3,5-trioxane dissociation. Molecular geometries and frequencies of all species were obtained at the B3LYP/cc-pVTZ, MP2/cc-pVTZ, and MP2/aug-cc-pVDZ levels of theory, whereas the single-point energies of the stationary points were calculated using coupled cluster with single and double excitations including the perturbative treatment of triple excitation (CCSD(T)) level of theory. It was found that the dissociation occurs via a concerted mechanism requiring an energy barrier of 48.3 kcal/mol to be overcome. The new experimental data and theoretical calculations serve as a validation and extension of kinetic data published earlier by other groups. Calculated values for the pressure limiting rate coefficient can be expressed as log10 k∞ (s(-1)) = [15.84 - (49.54 (kcal/mol)/2.3RT)] (500-1400 K).

A detailed theoretical analysis of the reaction of atomic bromine with tetrahydropyran (THP, C5H10O) was performed using several ab initio methods and statistical rate theory calculations. Initial geometries of all species involved in the potential energy surface of the title reaction were obtained at the B3LYP/cc-pVTZ level of theory. These molecular geometries were reoptimized using three different meta-generalized gradient approximation (meta-GGA) functionals. Single-point energies of the stationary points were obtained by employing the coupled-cluster with single and double excitations (CCSD) and fourth-order Møller-Plesset (MP4 SDQ) levels of theory. The computed CCSD and MP4(SDQ) energies for optimized structures at various DFT functionals were found to be consistent within 2 kJ mol(-1). For a more accurate energetic description, single-point calculations at the CCSD(T)/CBS level of theory were performed for the minimum structures and transition states optimized at the B3LYP/cc-pVTZ level of theory. Similar to other ether + Br reactions, it was found that the tetrahydropyran + Br reaction proceeds in an overall endothermic addition-elimination mechanism via a number of intermediates. However, the reactivity of various ethers with atomic bromine was found to vary substantially. In contrast with the 1,4-dioxane + Br reaction, the chair form of the addition complex (c-C5H10O-Br) for THP + Br does not need to undergo ring inversion to form a boat conformer (b-C4H8O2-Br) before the intramolecular H-shift can occur to eventually release HBr. Instead, a direct, yet more favorable route was mapped out on the potential energy surface of the THP + Br reaction. The rate coefficients for all relevant steps involved in the reaction mechanism were computed using the energetics of coupled cluster calculations. On the basis of the results of the CCSD(T)/CBS//B3LYP/cc-pVTZ level of theory, the calculated overall rate coefficients can be expressed as kov.,calc.(T) = 4.60 × 10(-10) exp[-20.4 kJ mol(-1)/(RT)] cm(3) molecule(-1) s(-1) for the temperature range of 273-393 K. The calculated values are found to be in excellent agreement with the experimental data published previously.

Radiation therapy for cancer patients works by ionizing damage to nuclear DNA, primarily by creating double-strand breaks (DSB). A major shortcoming of traditional radiation therapy is the set of side effect associated with its long-range interaction with nearby tissues. Low-energy Auger electrons have the advantage of an extremely short effective range, minimizing damage to healthy tissue. Consequently, the isotope ⁹⁹mTc, an Auger electron source, is currently being studied for its beneficial potential in cancer treatment. We examined the dose effect of a pyrene derivative ⁹⁹mTc complex on plasmid DNA by using gel electrophoresis in both aqueous and methanol solutions. In aqueous solutions, the average yield per decay for double-strand breaks is 0.011±0.005 at low dose range, decreasing to 0.0005±0.0003 in the presence of 1 M dimethyl sulfoxide (DMSO). The apparent yield per decay for single-strand breaks (SSB) is 0.04±0.02, decreasing to approximately a fifth with 1 M DMSO. In methanol, the average yield per decay of DSB is 0.54±0.06 and drops to undetectable levels in 2 M DMSO. The SSB yield per decay is 7.2±0.2, changing to 0.4±0.2 in the presence of 2 M DMSO. The 95% decrease in the yield of DSB in DMSO indicates that the main mechanism for DSB formation is through indirect effect, possibly by cooperative binding or clustering of intercalators. In the presence of non-radioactive ligands at a near saturation concentration, where radioactive Tc compounds do not form large clusters, the yield of SSB stays the same while the yield of DSB decreases to the value in DMSO. DSBs generated by ⁹⁹mTc conjugated to intercalators are primarily caused by indirect effects through clustering.

The oxidative dehydrogenation (ODH) reaction of propane was investigated at temperatures between 923 and 1023 K using either O2 or O2/H2S mixture as oxidant. GC analysis of the product mixtures showed that ethylene was the major olefin product in the conventional ODH reaction whereas propylene became dominant when H2S was included in the feed gas. With an oxygen-rich feed (4:2:2 C3H8:O2:H2S), ∼ 70% propane conversion, and ∼ 50% propylene selectivity could be achieved at 1023 K, a level of performance comparable to that for the ODH reaction employing reducible solid oxide catalysts. Theoretical calculations utilizing CBS-QB3 method were also conducted to explore the causes of the enhanced propylene yield and selectivity of the H2S-assisted ODH reaction. It was found that the increased propane conversion was due to a large enthalpy gain from the in situ formation of S2 that compensated for the high energy cost of hydrogen abstraction by SH and S2H. Also, the promoted propylene selectivity was attributed to the instability of the sulfur-containing products, which made the reaction route to propylene the most thermodynamically favored.

PEG-modified gold nanoparticles (PEG-modified GNs) with diameters of 40 nm and 70 nm were prepared to elucidate the effect of extent of PEG (M.W. 5000) grafting and particle size on tumor accumulation and cellular uptake. Flow cytometry reveals that cellular uptake is strongly related to the size of PEG-modified GNs, rather than the extent of PEG-5K grafting level. Cytotoxicity analysis based on the intracellular release of drugs showed that the 70 nm PEG-modified GNs have the higher cytotoxicity, beccause of their greater cellular uptake. Also, particle size, rather than PEG-5K grafting level affects tumor accumulation. However, PEG-5K grafting level significantly affects the accumulation of particles in the liver and spleen. This finding is important in determining the proper PEG-5K grafting level and particle size for designing nano-medicines., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Hydrogels have been developed as artificial extracellular matrixes (ECMs) to mimic native tissue microenvironments for various applications. Unfortunately, poly(N-isopropylacrylamide) (PNIPAAM)-based hydrogels are not suitable for cell culturing and cell sheet preparation. Carbon nanotubes (CNTs), with their mechanical strength and electrical conductivity, have been considered as additives to increase the applicability of hydrogels to cell encapsulation and advance cardiac electrophysiological functions. A simple method for fabrication of PNIPAAM hydrogels interpenetrated with multiwalled CNTs (MWCNTs) as substrates for cell sheet preparation is reported. The results demonstrate that PNIPAAM hydrogels with interpenetrating MWCNTs still exhibit thermosensitive behavior. It is also found that epithelial Madin-Darby canine kidney (MDCK) cells can only attach and proliferate on MWCNT-interpenetrated PNIPAAM hydrogels. Furthermore, the PNIPAAM hydrogels with MWCNTs possess higher elastic moduli and hydrophobicities than those without MWCNTs, suggesting these two characteristics are necessary for the cells to attach to the hydrogel surfaces. Moreover, cell sheets can only be harvested from PNIPAAM hydrogels with MWCNTs because of their high ratio of cell attachment. Thus, this simple method provides sufficient mechanical strength to PNIPAAM hydrogels so that anchorage-dependent cells can be cultivated and provides a superior system for preparing cell sheets., (Crown Copyright © 2013. Published by Elsevier Ltd. All rights reserved.)

In this study, lactoferrin-conjugated PEGylated liposomes (PL), a potential drug carrier for brain delivery, was loaded with radioisotope complex, 99mTc labeled N,N-bis(2-mercaptoethyl)-N',N'-diethylethylenediamine (99mTc-BMEDA) for in vitro and in vivo evaluations. The hydrophilicity of liposomes was enhanced by PEGylation which was not an ideal brain delivery system for crossing the blood brain barrier (BBB). With the modification of a brain-targeting ligand, lactoferrin (Lf), the PEGylated liposome (PL) might become a potential brain delivery vehicle. In order to test the hypothesis in vitro and in vivo, 99mTc-BMEDA was loaded into the liposomes as a reporter with or without Lf-conjugation. The mouse brain endothelia cell line, bEnd.3 cells, was cultured to investigate the potential uptake of liposomes in vitro. The in vivo uptake by the mouse brain of the liposomes was detected by tissue biodistribution study. The results indicated that Lf-conjugated PEGylated liposome showed more than three times better uptake efficiency in vitro and two-fold higher of brain uptake in vivo than PEGlyated liposome. With the success of loading the potential Single Photon Emission Tomography (SPECT) imaging probe, 99mTc-BMEDA, Lf-PL might serve as a promising brain delivery system for loading diagnostics or therapeutics of various brain disorders.

Purpose: The aim of the study is to assess the feasibility of imaging specific activity of myeloperoxidase (MPO), a leukocyte-derived enzyme with important role in atherosclerosis, by SPECT/CT using a novel radiotracer, (111)In-bis-5-hydroxytryptamide-diethylenetriamine-pentaacetate ((111)In-bis-5HT-DTPA)., Methods: Bis-5HT-DTPA was synthesized. Oligomerization of bis-5HT-DTPA in the presence of MPO/H(2)O(2) was studied and confirmed using MALDI-TOF. Apolipoprotein E knockout (ApoE KO) mice was used as an atherosclerosis-prone rodent model. Biodistribution assay and micro SPECT/CT imaging were carried out to prove the atherosclerosis targeting of (111)In-bis-5HT-DTPA in the ApoE KO mice., Results: MALDI-TOF spectrum showed that the 5HT base agent can self oligomerize after activating by MPO. From the biodistribution study, (111)In-bis-5HT-DTPA was quantified to be retained markedly higher while eliminated much slower in the aortas of the ApoE KO mice than that of the wild type (WT) mice within 1 h post-injection. The nuclear imaging showed significantly higher uptake in the aorta of the ApoE KO mice than that of the WT mice at least within 2 h post-injection., Conclusion: This study described the pharmacokinetics and biodistribution of (111)In-bis-5HT-DTPA in ApoE KO mice and validated its utilization for early detection of atherosclerotic marker, MPO, in the aortic wall of atherosclerosis-prone rodent model.

Background: Integrins are a family of transmembrane adhesion proteins that mediate cell adhesion and intracellular signaling. Integrin-αvβ3 is expressed on the surface of human glioblastoma cells, and can be further induced by chemical stress. The Arg-Gly-Asp (RGD) motif-containing peptides are specifically bound to integrin-αvβ3, and to inhibit neovasculature underlying competition to normal extracellular matrix proteins. This study employed two types of RGD peptides, cyclic RGD (c(RGDyK)) and bi-cyclic RGD (E[c(RGDyK)](2)) peptide, to human glioblastoma U87MG cells with combination of low dose Paclitaxel (PTX) pre-treatment to augment therapeutic activity for RGD peptide-induced apoptosis., Principal Findings: Human glioblastoma U87MG cells were treated with RGD peptides in the absence or presence of initial exposure to low-dose 10 nM PTX. Results showed that integrin-αvβ3 expressing on the surface of U87MG cells was induced by 10 nM PTX pre-treatment for 12 hrs. Additionally, the U87MG cells pre-treated with PTX and followed by RGD peptides exhibited greater expression of caspases-3, -8 and -9 than those merely treated with single agent of PTX or RGD peptide. Furthermore, the caspase-3, -8 and -9 inhibitor presented significant protection against E[c(RGDyK)](2) peptide induced U87MG programmed cell death. The increased expression of PTX-induced integrin-αvβ3 was correlated with the enhanced apoptosis in U87MG cells., Conclusions: This study provides a novel concept of targeting integrin-αvβ3 with RGD peptides in combination with low-dose PTX pre-treatment to improve efficiency in human glioblastoma treatment.

Target geometry for mitigating phagocytosis has garnered considerable attention recently in the drug delivery field. This study examined nanoparticles (NPs) with same volume but different shapes, namely, spherical NPs (SNPs) and hexagonal nanoprisms (HNPs), and analyzed their behaviors in vitro and in vivo. These NPs were constructed with a multifunctional block copolymer component, mPEG-b-P(HEMA-co-histidine-PLA). Geometry of SNPs and HNPs was controlled by adjusting copolymer properties and particle size was controlled by adjusting formulation parameters. Nanoparticle morphology had no effect in mitigating phagocytosis when NP size was 70 nm; however, morphology had a significant effect when NP size was 120 nm. The radioactivity-time curves for (99m)Tc-labeled NPs, fitted by the two-compartment pharmacokinetic model, show that the prolonged plasma distribution half-life of HNPs is indicative in the bloodstream. The in vitro and in vivo studies reveal that dual stealth characteristics, pegylation and hexagonal prism structure, of nanocarriers can be adopted in clinical application for safe and efficient delivery of cancer therapy., (Crown Copyright © 2011. Published by Elsevier B.V. All rights reserved.)

The rate coefficients for the reaction of 1,4-dioxane with atomic chlorine were measured from T = 292-360 K using the relative rate method. The reference reactant was isobutane and the experiments were made in argon with atomic chlorine produced by photolysis of small concentrations of Cl2. The rate coefficients were put on an absolute basis by using the published temperature dependence of the absolute rate coefficients for the reference reaction. The rate coefficients for the reaction of Cl with 1,4-dioxane were found to be independent of total pressure from p = 290 to 782 Torr. The experimentally measured rate coefficients showed a weak temperature dependence, given by k(exp)(T) = (8.4(-2.3)(+3.1)) × 10(-10) exp(-(470 ± 110)/(T/K)) cm3 molecule (-1) s(-1). The experimental results are rationalized in terms of statistical rate theory on the basis of molecular data obtained from quantum-chemical calculations. Molecular geometries and frequencies were obtained from MP2/aug-cc-pVDZ calculations, while single-point energies of the stationary points were computed at CCSD(T) level of theory. The calculations indicate that the reaction proceeds by an overall exothermic addition-elimination mechanism via two intermediates, where the rate-determining step is the initial barrier-less association reaction between the chlorine atom and the chair conformer of 1,4-dioxane. This is in contrast to the Br plus 1,4-dioxane reaction studied earlier, where the rate-determining step is a chair-to-boat conformational change of the bromine-dioxane adduct, which is necessary for this reaction to proceed. The remarkable difference in the kinetic behavior of the reactions of 1,4-dioxane with these two halogen atoms can be consistently explained by this change in the reaction mechanism.

The objective of this study was to investigate the cytotoxicity of the ethanolic extract of mycelia from Ganoderma lucidum (EMG) cultivated in a medium containing leguminous plants Glycine max (L.) Merr. and Astragalus membranaceus on human hepatocellular carcinoma cells (Hep 3B) and to isolate the active components from EMG. The results indicated that EMG induced cytotoxicity in a dose- and time-dependent manner, and the cells treated with EMG for 24, 48, and 72 h had IC(50) values of 156.8, 89.9, and 70.1 microg/mL, respectively. Furthermore, EMG was fractionated into seven fractions (F1-F7). We found that F5 and F6 had higher growth inhibitory effects on Hep 3B cells than the other fractions, and F6 possessed enough amounts (about 2.1 g) to carry out a more detailed study. F6 caused a sub-G1 peak rise and DNA fragmentation in Hep 3B cells and was further separated by high-performance liquid chromatography to obtain two active compounds, 9,11-dehydroergosterol peroxide [9(11)-DHEP] (compound 1) and ergosterol peroxide (EP) (compound 2). The IC(50) values of 9(11)-DHEP and EP based on the cell viability of Hep 3B were 16.7 and 19.4 microg/mL, respectively.

Introduction: Trastuzumab (Herceptin), a humanized IgG1 monoclonal antibody directed against the extracellular domain of the HER2 protein, acts as an immunotherapeutic agent for HER2-overexpressing human breast cancers. Radiolabeled trastuzumab with beta- or alpha emitters can be used as radioimmunotherapeutic agent for the similar purpose but with additional radiation effect., Methods: In this study, trastuzumab was labeled with (188)Re for radioimmunotherapy of HER2/neu-positive breast cancer. (188)Re(I)-tricarbonyl ion, [(188)Re(OH(2))(3)(CO)(3)](+), was employed as a precursor for directly labeling the monoclonal antibody with (188)Re. The immunoreactivity of (188)Re(I)-trastuzumab was estimated by competition receptor-binding assay using HER2/neu-overexpressive BT-474 human breast cancer cells. The localization properties of (188)Re(I)-trastuzumab within both tumor and normal tissues of athymic mice bearing BT-474 human breast cancer xenografts (HER2/neu-overexpressive) and similar mice bearing MCF-7 human breast cancer xenografts (HER2/neu-low expressive) were investigated., Results: When incubated with human serum albumin and histidine at 25 degrees C, (188)Re(I)-trastuzumab was found to be stable within 24 h. The IC(50) of (188)Re(I)-trastuzumab was found to be 22.63+/-4.57 nM. (188)Re(I)-trastuzumab was shown to accumulate specifically in BT-474 tumor tissue in in vivo biodistribution studies. By microSPECT/CT, the image of (188)Re localized BT-474 tumor was clearly visualized within 24 h. In contrast, (188)Re(I)-trastuzumab uptake in HER2-low-expressing MCF-7 tumor was minimal, and the (188)Re image at the localization of the tumor was dim., Conclusion: These results reveal that (188)Re(I)-trastuzumab could be an appropriate radioimmunotherapeutic agent for the treatment of HER2/neu-overexpressing cancers.

By simply incubating Herceptin (trastuzumab) with [99m Tc(CO)3(OH2)3]+ ion in saline, a significant yield of 99m Tc-labeled trastuzumab was found to be achievable. The effective labeling may be based on that trastuzumab is inherent with endogenous histidine group to which 99m Tc(I) tricarbonyl ion can be strongly bound. For practical 99m Tc labeling processing, trastuzumab was purified beforehand from the commercial product, Herceptin (Genentech) via size exclusion chromatography to remove the excipient, alpha-histidine and a high-labeled yield could be obtained by incubating the purified trastuzumab with [99m Tc(CO)3(OH2)3]+. Retention of bioactivity of the 99m Tc(I)-labeled trastuzumab was validated using a cell binding test.

A cell survival assay of the four arabinosyl uridine analogs with functionalities of 5-fluoro, 5-fluorovinyl, 5-iodo, and 5-iodovinyl as potential positron-emitter tagged probe for monitoring cancer gene therapy were performed. Cytotoxicities of 5-fluoro-, 5-iodo-, 5-fluorovinyl, and 5-iodovinyl arabinosyl uridines against SR-39 thymidine kinase transfected murine prostate cancer cells have been evaluated using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. None of them showed significant bioactivity. A syn conformation derived from intra-hydrogen bonding was suggested for the unfavorable interaction and diminished bioactivity.

Sediments as a tool for monitoring contamination by heavy metals in the environment has long been considered. It is therefore a necessity to produce reliable data for such purposes. Microwave-assisted acid dissolution has proved to be a suitable method for digesting complex matrices, such as sediments. However, due to the infancy of the technique, the procedures are numerous and varied in both the reagents used and microwave conditions. In this study, the efficiency of two recommended acid mixtures, a HNO3-HF mixture and an aqua regia-HF mixture, under the same microwave digestion conditions were compared using certified reference materials. It was observed that the HNO3-HF mixture showed better efficiency than the aqua regia-HF mixture in most of the heavy metals analyzed in all certified reference materials used.

Oxorhenium(V) complexes of two thioether amines have been synthesized and characterized spectroscopically. The five-coordinated ReO(3+) moiety is satisfied by the amine compounds behaving as dibasic tetradentate (NSSN) ligands and a chloride ion. The radioanalytical studies of their technetium analogs have been carried out at different pH values in the range 3-10. The solvent extraction, TLC, electrophoresis and HPLC studies using (99m)Tc radiotracer indicate that the labeled complex with the TcO(3+) core is similar to the rhenium complex with the ReO(3+) core.

The title compound, C(10)H(18)N(2)S(2), acts as an important precursor for the synthesis of the pharmaceutically important diaminedithiol ligand system. The molecule has a local twofold axis and the arrangement of the S(2)N(2) donor atoms in the macrocycle is anticlinal.

An activity-directed fractionation and purification process was used to identify the antioxidative components of adlay hulls. Hulls of adlay (Coix lachryma-jobi L. var. ma-yuen Stapf) were extracted with methanol and then separated into water, 1-butanol, ethyl acetate, and hexane fractions. The 1-butanol-soluble fraction exhibited greater capacity to scavenge 2,2'-diphenyl-1-picrylhydrazyl (DPPH) radicals when compared with fractions soluble in water, ethyl acetate, and hexane phases. The 1-butanol fraction was then subjected to separation and purification using Diaion HP-20 chromatography, silica gel chromatography, and HPLC. Six compounds showing strong antioxidant activity were identified by spectroscopic methods ((1)H NMR, (13)C NMR, IR, and MS) and by comparison with authentic samples to be coniferyl alcohol (1), syringic acid (2), ferulic acid (3), syringaresinol (4), 4-ketopinoresinol (5), and a new lignan, mayuenolide (6).

The title compound, [Re(2)O(3)(C(19)H(20)N(2)O(2))(2)], is a hexacoordinate complex containing an [Re(2)O(3)](4+) core with a linear O=Re-O-Re=O bridge. The distorted octahedral coordination of the Re(V) atom is achieved by an N(2)O(2) donor set from the tetradentate imine-phenol ligand. The overall charge of the compound is neutral due to deprotonation of the phenol groups, and the terminating and bridging O atoms. The Re=O and Re-O bond distances of the [Re(2)O(3)](4+) core are 1.699 (4) and 1.911 (1) A, respectively. The Re-O and Re-N bond distances of the equatorial plane are in the ranges 2.024 (4)-2.013 (4) and 2.128 (5)-2.120 (5) A, respectively.

A rhenium(V) complex with 2-(2-pyridylmethylthio)-aniline has been synthesized and characterized by UV-VIS absorption, IR, MS and X-ray crystallography. The complex contains a ReO3+ moiety and the amine compound, acting as a monobasic tridentate(NSN) ligand. The chemical behavior of its technetium analog has been studied radioanalytically by solvent extraction, TLC, electrophoresis and HPLC using 99mTc tracer.

A convenient four-step procedure was developed to prepare the novel monoprotic tetradentate ligands N-(2-hydroxy)benzyl-N-methyl-N'-(2-pyridyl)methyl-1,3-propanediamine (Hpamap) and N-(2-hydroxy)benzyl-N'-methyl-N'-(2-pyridyl)methyl-1,3-propanediamine (Hpmaap), which provide an N(3)O metal coordination sphere. A mononuclear copper(II) complex, [Cu(pamap)Cl] (A), was obtained by reaction of Hpamap with CuCl(2).2H(2)O. The binuclear copper(II) complexes [Cu(pamap)](2)(BF(4))(2) (B) and [Cu(pmaap)](2)(BF(4))(2) (C) were obtained when these ligands were reacted with Cu(II) in the presence of the noncoordinating BF(4)(-) anion. Reaction of nickel(II) with the Hpamap ligand generated the binuclear Ni(II) complex [Ni(2)(pamap)(2)(NO(3))]NO(3) (D). The crystal of A (C(17)H(22)ClCuN(3)O) is orthorhombic Pbca (No. 61), a = 11.837(4) Å, b = 15.648(5) Å, c = 11.002(11) Å, Z = 8; that of B (C(34)H(44)B(2)Cu(2)F(8)N(6)O(2)) is triclinic P&onemacr; (No. 2), a = 9.147(0) Å, b = 10.375(0) Å, c = 10.535(1) Å, alpha = 107.20(0) degrees, beta = 91.19(0) degrees, gamma = 105.05(0) degrees, Z = 1; that of C (C(34)H(44)B(2)Cu(2)F(8)N(6)O(2)) is monoclinic P2(1)/c (No. 14), a = 9.158(2) Å, b = 10.714(2) Å, c = 19.085(4) Å, beta = 90.58(2) degrees, Z = 4; and that of D (C(34)H(44)N(8)NiO(8)) is monoclinic C2/c (No. 15), a = 13.849(0) Å, b = 13.609(0) Å, c = 19.558(1) Å, beta = 92.34 (0) degrees, Z = 4. The copper atoms of all three complexes are five-coordinate in the solid state, assuming the geometry of a distorted square pyramid with the deprotonated tetradentate ligand in the basal plane. The mononuclear complex A has a chloride ligand in the axial position, while each copper center in the binuclear complexes B and C has, in the axial position, a bridging phenolate O donor from the other unit of the dimer. Each nickel center in the binuclear complex D is six-coordinate, with the pseudo-octahedron formed by a deprotonated tetradentate ligand, a bridging nitrato oxygen atom, and a bridging phenoxy donor from the tetradentate ligand bound to the second nickel center.

The aim of this study was to evaluate 99Tcm-labelled phenylene imine phenol (99Tcm-Ph(IP)2) as a leukocyte label. The 99Tcm complex exhibits a neutral lipophilic character. The radiochemical purity of the 99Tcm-Ph(IP)2 was high both soon after labelling (94.5 +/- 1.2%, n = 7) and at 24 h (89.7 +/- 1.0%, n = 7). In a comparative study, the labelling efficiency of 99Tcm-Ph(IP)2 and 99Tcm-hexamethylpropylene amine oxime (99Tcm-HMPAO) labelled leukocytes was 47.1 +/- 3.5% (n = 7) and 58.5 +/- 7.0% (n = 7) respectively. The viability of labelled leukocytes was greater than 90% for both 99Tcm complexes. The elution ratios of labelled leukocytes with 99Tcm-Ph(IP)2 at 1, 4 and 24 h were 10.1 +/- 3.5%, 17.2 +/- 2.7% and 23.0 +/- 2.4% (n = 7) respectively, while those with 99Tcm-HMPAO were 10.8 +/- 1.2%, 18.3 +/- 2.0% and 37.0 +/- 3.7% (n = 7) respectively. In conclusion, 99Tcm-Ph(IP)2, with good stability and significant leukocyte labelling efficiency, is a potential leukocyte label.

Synthesis and characterization of a new pentadentate amine-oxime ligand, 3,3,11,11-tetramethyl-7-benzyl-4,7,10-triazatridecane-2,12- dionedioxime is reported. The technetium complexes of this ligand was formed by direct reduction of TcO4- in the presence of the ligand at pH 9. The radiochemical yield of the complex is greater than 95% with 10(-7) M 99TcO4-. The technetium complex formed is a positively charged chelate that elutes as a single peak on a reverse phase HPLC. Biodistribution studies in rats showed an uptake of 1.4% of the injected activity at 15 sec p.i. in heart, but the wash-out was fast. The high radiochemical purity and stability of this 99mTc-chelate indicate that these types of chelates hold potential for development of bifunctional chelating agents (BFCAs).

An attempt was made to use 99Tcm-ethyl cysteinate dimer (99Tcm-ECD) (Neurolite, Du Pont Merck, N. Billerica, MA) to label leukocytes. The radiochemical purity of 99Tcm-ECD, labelling efficiency of leukocytes, cell viability of labelled leukocytes and stability of 99Tcm-ECD-labelled leukocytes were calculated. Compared with the commercial cell-labelling agent, 99Tcm-hexamethylpropyleneamine oxime (99Tcm-HMPAO): (1) the radiochemical purity of 99Tcm-ECD was higher than that of 99Tcm-HMPAO and at immediately, 1, 2, 4, 6, 8 and 24 h after 99Tcm labelling; (2) the labelling efficiency of 99Tcm-ECD-labelled leukocytes was lower than that of 99Tcm-HMPAO; (3) the viability of the labelled white blood cells (WBC) was high for both agents; and (4) the stability of 99Tcm-ECD-labelled leukocytes was worse than that of 99Tcm-HMPAO at 1, 2, 4, 6, 8 and 24 h. It is concluded that although 99Tcm-ECD is more stable than 99Tcm-HMPAO, because of the lower labelling efficiency and power stability of 99Tcm-ECD-labelled leukocytes, 99Tcm-ECD is not a good choice as a leukocyte-labelling agent to replace commercial 99Tcm-HMPAO.

99mTc-diamine-diphenol chelates are neutral lipophilic chelates exhibiting good stability in aqueous solutions. The cell labeling and biolocalization properties of four different 99mTc-amine-phenol complexes were determined. All four chelates readily labeled leukocytes and RCBs in high yields. Even though 99mTc was retained by the cells, the elution rate of 99mTc from the labeled cells in plasma at 37 degrees C was unacceptably high for potential utility in scintigraphic imaging. The uptake of 99mTc in brain or heart following i.v. injection of the chelates in rats was low and clearance of activity from the blood was slow.

The conjugation of a complex formed by reacting RhCl3 with cysteine to human serum albumin has been investigated. Approximately 50% of the rhodium (labeled with 105Rh) was converted to the complex. Conjugation of the complex to HSA via the ECDI method resulted in yields of approximately 40% of the total rhodium or approximately 80% of the Rh-cysteine complex. No conjugation was observed in the absence of the ECDI. At approximately equal molar concentrations of rhodium and HSA, an average of approximately 0.4 rhodium atoms per HSA molecule was achieved.

A bifunctional ligand 4-(p-aminobenzyl)-diethylenetriamine has been synthesized. 105Rh complexes with this ligand were prepared with an overall yield of 79% at pH 9.0 in bicarbonate buffer. The preformed complex was converted to the isothiocyanate derivative using thiophosgene. Conjugation yields of 75% with IgG and 85% with HSA could be obtained for a 4 h conjugation reaction. Affinity chromatography of human-IgG coupled to the rhodium complex in an anti-human IgG agarose gel indicated no denaturation of the labeled protein. The procedure reported here can be adapted for the preparation of 105Rh-labeled antibodies for therapeutic applications.

Labeling of hematoporphyrin with 105Rh at stoichiometric concentrations is described. Labeling efficiencies of up to 93% could be obtained at pH 9.0 in bicarbonate buffer. Solvent extraction of 105Rh-hematoporphyrin into methyl isobutyl ketone was used to estimate the complex yield. The complex showed high stability and no loss of 105Rh was seen throughout the 6 days of study. 105Rh-hematoporphyrin when incubated with human gamma globulin was seen to be quantitatively bound to the protein. This procedure may be used for labeling monoclonal antibodies with 105Rh for therapeutic applications.