Cancer therapy

In recent years a growing recognition that molecularly-targeted therapies face formidable obstacles has revived interest in more generic tumor cell phenotypes that could be exploited for therapy. Two recent reports demonstrate that cancer cell survival is critically dependent on the activity of MTH1, a nucleotide pyrophosphatase that converts the oxidized nucleotides 8-oxo-dGTP and 2-OH-dATP to the corresponding monophosphates, thus preventing their incorporation into genomic DNA. Tumor cells frequently overexpress MTH1, probably because malignant transformation creates oxidative stress that renders the nucleotide pool highly vulnerable to oxidation. As a result, MTH1 inhibition in cancer cells results in accumulation and incorporation of 8-oxo-dGTP and 2-OH-dATP into DNA, leading to DNA damage and cell death. This toxic effect is highly cancer cell-specific, as MTH1 is generally dispensable for the survival of normal, untransformed cells. Importantly, MTH1 proves to be a “druggable” enzyme that can be inhibited both by an existing protein kinase inhibitor drug, crizotinib, and by novel compounds identified through screening. Inhibition of MTH1 leading to toxic accumulation of oxidized nucleotides specifically in tumor cells therefore represents an example of a “non-personalised” approach to cancer therapy.     

Macrocystis (Laminariales,Phaeophyceae) in South Africa: potential for cultivation through holdfast fragmentation and use as feed for the aquacultured abalone,Haliotis midae

Journal of Applied Phycology - Macrocystis pyrifera (Linnaeus) C. Agardh, known in some regions as ‘giant kelp’, occurs along temperate coastal rocky reefs throughout the southern...     

ADP-ribosylation factor is a subunit of the coat of Golgi-derived COP-coated vesicles: a novel role for a GTP-binding protein.

ADP-ribosylation factor (ARF) is an abundant and highly conserved low molecular weight GTP-binding protein that was originally identified as a key element required for the action of cholera toxin in mammalian cells, but whose physiological role is unknown. We report that ARF family proteins are highly concentrated in non-clathrin-coated transport vesicles and are coat proteins. About three copies of ARF are present on the outside of coated vesicles per alpha-COP (and thus per coatomer). ARF is highly enriched in coated vesicles as compared with parental Golgi cisternae, as shown both by biochemical and morphological methods, and ARF is removed from transport vesicles through uncoating during transport. Furthermore, ARF binds to Golgi cisternae in a GTP-dependent manner independently of coated vesicle budding. These observations strongly suggest a new role for GTP-binding proteins: ARF proteins may modulate vesicle budding and uncoating through controlled GTP hydrolysis.     

Interindividual variation in the levels of certain urinary polycyclic aromatic hydrocarbon metabolites following medicinal exposure to coal tar ointment

Determination of human exposure to polycyclic aromatic hydrocarbons PAHs is challenging because they are so broadly distributed in the environment and often difficult to quantitate using questionnaire methods. To enhance the ability to non invasively evaluate markers of both internal dose and biologically effective dose we have developed methods for the identification and quantitation of 1 hydroxypyrene-glucuronide and r 7, t 8, t 9, c 10 tetrahydroxy 7,8,9,10 tetrahydrobenzo a pyrene BP 7,10 8,9 tetrol in human urine. In the current study we applied these assays to urine samples collected from 43 hospitalized psoriasis patients treated with coal tar medication and 39 non treated volunteer controls. BP 7,10 8,9 tetrol was detected in 20 of 43 47 patients, ranging from 1 not detected to 124 fmol mumol-1 creatinine. In contrast, BP 7,10 8,9 tetrol was detected in only 4 of 39 10 controls, range 1 to 20.6 fmol mumol-1 creatinine p = 0.0006, Wilcoxon rank sum test . A second, more polar PAH metabolite, identified as 1 hydroxypyrene-glucuronide, was present in all urine samples. Mean 1 hydroxypyreneglucuronide levels were 40.96 72.62 pmol mumol-1 creatinine in patients and 0.38 0.32 pmol mumol-1 creatinine in control subjects p 0.0001 . The ratio of urinary levels of BP 7,10 8,9 tetrol to 1 hydroxypyrene-glucuronide was examined in the coal tar treated patients. This ratio was found to vary by approximately 6000 fold. This parameter cannot be explained by measurement error because the coefficients of variation for these assays are only 12 and 10 respectively, nor can it be explained by use of different coal tar products. These results provide further evidence that substantial interindividual variation in activation of benzo a pyrene and other PAHs exists, which may have implications for disease risk.     

Carbon-13 nuclear magnetic resonance studies of myocardial glycogen metabolism in live guinea pigs

Myocardial glycogen metabolism was studied in live guinea pigs by 13C NMR at 20.19 MHz. Open-chest surgery was used to expose the heart, which was then positioned within a solenoidal radio frequency coil for NMR measurements. The time course of myocardial glycogen synthesis during 1-h infusions of 0.5 g of D-[1-13C]glucose (and insulin) into the jugular vein was investigated. The possible turnover of the 13C-labeled glycogen was also studied in vivo by following the labeled glucose infusion with a similar infusion of unlabeled glucose. The degree of 13C enrichment of the C-1 glycogen carbons during these infusions was measured in heart extracts by 1H NMR at 360 MHz. High-quality proton-decoupled 13C NMR spectra of the labeled C-1 carbons of myocardial glycogen in vivo were obtained in 1 min of data accumulation. This time resolution allowed measurement of the time course of glycogenolysis of the 13C-labeled glycogen during anoxia by 13C NMR in vivo. With the solenoidal coil used for 13C NMR, the spin-lattice relaxation time of the labeled C-1 carbons of myocardial glycogen could be measured in vivo. For a comparison, spin-lattice relaxation times of heart glycogen were measured in vitro at 90.55 MHz. Natural abundance 13C NMR studies of the quantitative hydrolysis of extracted heart glycogen in vitro at 90.55 MHz showed that virtually all the carbons in heart glycogen contribute to the 13C NMR signals. The same result was obtained in 13C NMR studies of glycogen hydrolysis in excised guinea pig heart.     

Peptide-binding antibiotics: A solid-phase assay for screening libraries of vancomycin mimics for selective d-Ala-d-Ala binding

Due to recent increases of vancomycin-resistant bacterial infections, research on more effective antibiotics has intensified. As part of such research, screening of combinatorial libraries is a valuable approach for discovering new lead antibiotics. A screening method is presented here that uses a solid state library and a fluorescent label to detect RCO-d-Ala-d-Ala binding in aqueous media.