Multiarray cell stretching platform for high-magnification real-time imaging.

AIM: This article reports the development of a multiarray microchip with real-time imaging capability to apply mechanical strains onto monolayered cell cultures. MATERIALS & METHODS: Cells were cultured on an 8-µm thick membrane that was positioned in the microscope focal plane throughout the stretching process. Each stretching unit was assembled from three elastomeric layers and a glass coverslip. A programmable pneumatic control system was developed to actuate this platform. Multiple stretching experiments were conducted with various cell lines. RESULTS: The platform provides a maximum uniform strain of 69%. Acute and long-term cell morphological changes were observed. The supreme imaging capability was verified by real-time imaging of transfected COS-7 stretching and poststretching imaging of immunofluorescence-stained PTK2. CONCLUSION: The platform reported here is a powerful tool for studying mechanically induced physiological changes in cells. Such a device could be used in tissue regeneration for maintaining essential cell growth conditions.

Rhenium(I) tricarbonyl complexes of salicylaldehyde semicarbazones: synthesis, crystal structures and cytotoxicity.

A series of N,N-disubstituted salicylaldehyde semicarbazones (SSCs), HOC(6)H(4)CHN-NHCONR(2), and their rhenium(I) tricarbonyl complexes, [ReBr(CO)(3)(SSC)], have been synthesised and characterised by IR and (1)H NMR spectroscopy. Crystallographic analysis of the complex [ReBr(CO)(3)(H(2)Bu(2))] (H(2)Bu(2)=SSC where R=Bu(n)) showed that the SSC acts as a bidentate ligand via its imino nitrogen and carbonyl oxygen atoms. The [ReBr(CO)(3)(SSC)] complexes exhibit moderate to high cytotoxicities towards MOLT-4 cells (IC(50)=1-24µM, cf. 18µM for cisplatin), and the majority of them are virtually non-toxic against non-cancerous human fibroblasts. Apoptotic assays of [ReBr(CO)(3)(H(2)Bnz(2))] (Bnz=benzyl) revealed that it mediates cytotoxicity in MOLT-4 cells via apoptosis. The complex [ReBr(CO)(3)(H(2)Bnz(2))] reacts with guanosine by proton transfer from the phenolic OH group to N(7) of guanosine. In (CD(3))(2)SO, [ReBr(CO)(3)(H(2)Bnz(2))] undergoes facile conversion to the dimeric complex, [Re(CO)(3)(HBnz(2))](2), via bromide dissociation.

Rapid determination of vitamin B12 concentration with a chemiluminescence lab on a chip.

This paper reports a novel method for the rapid determination of vitamin B(12) concentration in a continuous-flow lab-on-a-chip system. This new method is based on luminol-peroxide chemiluminescence (CL) assays for the detection of cobalt(II) ions in vitamin B(12) molecules. The lab-on-a-chip device consisted of two passive micromixers acting as microreactors and a double spiral microchannel network serving as an optical detection region. This system could operate in two modes. In the first mode, samples are acidified and evaluated directly in the microchip. In the second mode, samples are treated externally by acidification prior to detection in the microchip. In the first mode, the linear range obtained was between 1.00 ng ml(-1) to 10 µg ml(-1), R(2) = 0.996, with a relative standard deviation (RSD) of 1.23 to 2.31% (n = 5) and a limit of detection (lod) of 0.368 pg ml(-1). The minimum sample volume required and the analytical time were 30 µl and 3.6 s, respectively. In the second mode, the linear range obtained was between 0.10 ng ml(-1) to 10 µg ml(-1), R(2) = 0.994, with the RSD of 0.90 to 2.32% (n = 6) and a lod of 0.576 pg ml(-1). The minimum sample and the analytical time required were 50 µl and 6 s, respectively. The lab on a chip working in mode II was successfully used for the determination of vitamin B(12) concentrations in nutritional supplemental tablets and hen egg yolks.

DNA binding and nucleolytic properties of Cu(ii) complexes of salicylaldehyde semicarbazones.

The copper(ii) complexes of two salicylaldehyde semicarbazones, HOC(6)H(4)CH[double bond, length as m-dash]N-NHCONR(2) [H(2)Bnz(2) (R = CH(2)Ph) and H(2)Bu(2) (R = Bu)], were evaluated for their DNA binding and cleavage properties by spectrophotometric DNA titration, ethidium bromide displacement assay and electrophoretic mobility shift assay. Results showed that the Cu(ii) complexes can bind to DNA via a partial intercalation mode with binding constants of 1.1 × 10(4) and 9.5 × 10(3) M(-1) for [Cu(HBnz(2))Cl] and [Cu(HBu(2))Cl], respectively. These complexes also cleave DNA in the presence of ascorbic acid, most likely through hydroxyl radicals that are generated via the reduction of a Cu(ii) to a Cu(i) species. The complexes show similar DNA cleavage activity, which is reflected in the similarity of their frontier molecular orbital energies calculated by density functional theory. These results are discussed in relation to the anticancer properties of the complexes.

Cytotoxic copper(II) salicylaldehyde semicarbazone complexes: mode of action and proteomic analysis.

The in vitro cytotoxic studies of a series of salicylaldehyde semicarbazones, HOC6H4CH=N-NHCONR2 (H2R2) and their Cu(II) complexes on a number of human tumor cell lines were conducted and it was observed that their cytotoxicities were enhanced following complexation to copper. These copper(II) complexes also demonstrated higher in vitro activities than the reference drug, cisplatin, on the tumor cell lines at micro molar range. Apoptotic assays and cell cycle analysis of the copper complexes, [Cu(HBnz2)Cl] and [Cu(HBu2)Cl] revealed that they mediated cytotoxicity in MOLT-4 cells via apoptosis. Further proteomic investigation of [Cu(HBnz2)Cl] and [Cu(HBu2)Cl] with respect to their protein expression profiles associated with their mode of action was conducted. By comparing the expression levels of 33 identified protein spots amongst the respective compound-treated profiles, we identified similarities in protein expression patterns between the two copper(II) complexes. The possible roles of the identified proteins in the execution of apoptosis by these copper(II) complexes are discussed.

Copper, gold and silver compounds as potential new anti-tumor metallodrugs.

Although platinum-based drugs such as cisplatin are powerful anticancer agents, they have undesirable side effects and are effective against only a few kinds of cancers. There is, therefore, a need for new drugs with an improved spectrum of efficacy and lower toxicity. Complexes of copper, gold and silver (coinage metals) are potential candidates to fulfill this need. The development of anticancer drugs based on these metals is currently a very active field. Considerable effort has also been put into elucidating the mechanisms of action of these complexes and optimizing their bioactivity through structural modification. In this review, we highlight recent developments in the design of coinage metal complexes with anti-tumor activity and discuss the emerging importance of quantitative structure-activity relationship methods in the study of anticancer metal complexes. Future work in this field, including likely coinage metal complexes that will attract attention, are proposed.

Hantupeptins B and C, cytotoxic cyclodepsipeptides from the marine cyanobacterium Lyngbya majuscula.

Hantupeptins B (2) and C (3) were isolated, along with the previously reported hantupeptin A (1), from the marine cyanobacterium, Lyngbya majuscula, collected from Pulau Hantu Besar, Singapore. Their structures were elucidated by interpretation of extensive 1D and 2D NMR spectroscopic data. Compounds 2 and 3 are cyclic depsipeptides consisting of five alpha-amino/hydroxy acid residues, including phenyllactic acid, proline, N-methyl-valine, valine, N-methyl-isoleucine, and a beta-hydroxy acid unit with different degrees of unsaturation at the terminal end of each molecule. The absolute configurations of the common amino acids and phenyllactic acid were determined by the advanced Marfey's and chiral HPLC analyses, respectively. The complete stereochemistry of 3-hydroxy-2-methyl-7-octynoic acid moiety in hantupeptin A was elucidated by a combination of homonuclear J-resolved 2D NMR experiments and by Mosher's method. Hantupeptins B and C showed moderate in vitro cytotoxicity when tested against MOLT-4 (leukemic) and MCF-7 (breast cancer) cell lines.

Synthesis and cytotoxic activities of chloropyridylimineplatinum(II) and chloropyridyliminecopper(II) surface-functionalized poly(amidoamine) dendrimers.

The preparations of novel platinum and copper metallodendrimers are reported. Surface modified first generation (G0) poly(amidoamine) (PAMAM) dendritic Schiff base, prepared via a condensation reaction was coordinated with platinum chloride and copper chloride yielding [G0-Py(4)-[PtCl(2)](4)] (4D) and [G0-Py(4)-[CuCl(2)](7)] (7E) respectively. These functionalized hyper-branched complexes were characterized by IR spectroscopy and CHN analysis. 4D was further characterized through (1)H and (13)C spectroscopy, while 7E was characterized using matrix-assisted laser desorption ionization time-of-flight (MALDI/TOF) Mass Spectrometer. The cytotoxic effects of the compounds against cells of neoplastic origin (MOLT-4, MCF-7) and cells of benign origin (Chang Liver) were studied. Their cytotoxicities were then compared to their mono-nuclear analogues, [(MeCONHCH(2)CH(2)NCHPy)(PtCl(2))] (1D) and [(MeCONHCH(2)CH(2)NCHPy)(CuCl(2))] (1E). The multi-nuclear complexes showed increased cytotoxic activities as compared to their respective mono-nuclear compounds. Most notably, significant inhibitions were observed for 7E on all cell lines, in which its IC(50) values were 11.1+/-0.6, 10.2+/-1.5 and 8.7+/-0.7microM against MOLT-4, MCF-7 and Chang Liver cells respectively. The multi-nuclear copper-based complexes (7E) are therefore most effective against a cancer cell line (MOLT-4) and a cisplatin-resistant cell line (MCF-7).

Hantupeptin A, a cytotoxic cyclic depsipeptide from a Singapore collection of Lyngbya majuscula.

Chemical investigation of the marine cyanobacterium Lyngbya majuscula from Pulau Hantu Besar, Singapore, has led to the isolation of a cyclodepsipeptide, hantupeptin A (1). The planar structure of 1 was assigned on the basis of extensive 1D and 2D NMR spectroscopic experiments. The absolute configuration of the amino and hydroxyl acid residues in the molecule was determined by application of the advanced Marfey method, chiral HPLC analysis, and Mosher's method. Hantupeptin A showed cytotoxicity to MOLT-4 leukemia cells and MCF-7 breast cancer cells with IC(50) values of 32 and 4.0 microM, respectively.

Synthesis, crystal structures and cytotoxicities of some transition metal complexes with N-[2-{(pyridin-2-ylmethylidene)amino}ethyl]acetamide.

The synthesis and spectroscopic (IR, (1)H and (13)C NMR) characterization of new complexes of Pt(II), Pd(II), Cu(II), and Hg(II) with the Schiff base ligand MeCONHCH(2)CH(2)N=CHPy (L) (Py=pyridine) are reported, together with studies on the cytotoxicities of these complexes, L and [ReBr(CO)(3)(L)] against human leukemia (MOLT-4), breast cancer (MCF-7) and Chang Liver (non-cancerous) cells. The crystal structures of [Pt(L)Cl(2)] (2), [Cu(L)Cl(2)] (4) and [Hg(L)Cl(2)](2) (5) are also reported. Of the complexes studied, [Cu(L)Cl(2)] (4) was identified as the most cytotoxic active derivative against cells of neoplastic origin (MOLT-4, and MCF-7), while having low toxicity on cells of benign origin (Chang Liver).