To better assess the potential biological consequences of diagnostic x-rays and selected gamma-emitting radioisotopes used in brachytherapy, we used the PENELOPE Monte Carlo radiation transport code to estimate the spectrum of initial electrons produced by photons in single cells and in an irradiation geometry similar to those used in cell culture experiments. We then combined estimates of the initial spectrum of electrons from PENELOPE with DNA damage yields for monoenergetic electrons from the fast Monte Carlo damage simulation (MCDS). The predicted absolute yields (Gbp(-1) Gy(-1)) and RBE values for single-strand break (SSB) and double-strand break (DSB) induction by 220 kVp x-rays are within 1% of the results from detailed track-structure simulations (Friedland et al 1999 Radiat. Environ. Biophys. 38 39). The measured RBE for DSB induction reported by Kühne et al (2005 Radiat. Res. 164 669) for gamma-rays from (60)Co and for 29 kVp x-rays with a 50 microm Rh (mammography) filter are in excellent agreement (1.15 versus 1.16). DSB yields predicted by the MCDS also agree to within 7% with the absolute DSB yields reported by de Lara et al (2001 Radiat. Res. 155 440) and Botchway et al (1997 Radiat. Res. 148 317) for the irradiation of V79 cells by low energy (<2 keV) characteristic x-rays. The predicted RBE for DSB induction by gamma-rays from bare (169)Yb and (131)Cs to (60)Co are 1.06 and 1.14, respectively. Tabulated RBE values for the single-cell and monolayer cell culture geometries differ by at most 15%. The proposed methodology is computationally efficient and may also be useful for the prediction of damage yields for mixtures of other types of charged particles, such as those found in proton therapy, space applications or internal dosimetry.

The Rana catesbeiana (bullfrog) ribonucleases, which belong to the RNase A superfamily, exert cytotoxicity toward tumor cells. RC-RNase, the most active among frog ribonucleases, has a unique base preference for pyrimidine-guanine rather than pyrimidine-adenine in RNase A. Residues of RC-RNase involved in base specificity and catalytic activity were determined by sitedirected mutagenesis, k cat /K m analysis toward dinucleotides, and cleavage site analysis of RNA substrate. The results show that Pyr-1 (N-terminal pyroglutamate), Lys-9, and Asn-38 along with His-10, Lys-35, and His-103 are involved in catalytic activity, whereas Pyr-1, Thr-39, Thr-70, Lys-95, and Glu-97 are involved in base specificity. The cytotoxicity of RC-RNase is correlated, but not proportional to, its catalytic activity. The crystal structure of the RC-RNase⅐d(ACGA) complex was determined at 1.80 Å resolution. Residues Lys-9, His-10, Lys-35, and His-103 interacted directly with catalytic phosphate at the P 1 site, and Lys-9 was stabilized by hydrogen bonds contributed by Pyr-1, Tyr-28, and Asn-38. Thr-70 acts as a hydrogen bond donor for cytosine through Thr-39 and determines B 1 base specificity. Interestingly, Pyr-1 along with Lys-95 and Glu-97 form four hydrogen bonds with guanine at B 2 site and determine B 2 base specificity.Ribonucleases are found widely within living organisms and are thought to play an important role in the metabolism of RNA. Recently, it has been shown that several members of the bovine ribonuclease superfamily exhibit biological functions in addition to intrinsic ribonucleolytic activities. Human eosinophil-derived neurotoxin and eosinophil cationic protein exert neurotoxicity (1) as well as antiparasitic activity (2), human angiogenin induces blood vessel formation (3), and frog ribonuclease exhibits antitumor activity (4, 5). Ribonucleolytic activity is essential for the biological functions of these proteins (6 -12).Bovine pancreatic ribonuclease, known as RNase A, in the ribonuclease superfamily is well characterized and is a valuable model for the study of structure-function relationships and protein refolding (13,14). It consists of 124 amino acid residues linked with four pairs of disulfide bridges and possesses a substrate preference for pyrimidine-adenosine in the RNA sequence but no cytotoxicity toward tumor cells. There are three subsites within RNase A molecule: the P 1 site, at which phosphodiester bond cleavage occurs; the B 1 site, for binding pyrimidine, which donates oxygen via its ribose to the scissile bond; and the B 2 site, for binding the adenine ring on the opposite site of the scissile bond. Three amino acid residues, His-12, Lys-41, and His-119, at the P 1 site are involved in catalytic activity. Four amino acid residues, Thr-45, Asp-83, Phe-120, and Ser-123, at the B 1 site are involved in the binding of the 5Ј-ribonucleoside, pyrimidine, whereas two residues, Asn-71 and Glu-111, at the B 2 site are involved in the binding of the 3Ј-ribonucleoside, adenosine (14 -18).A new group of ribon...

CC chemokine receptors 2 (CCR2) and 5 (CCR5) are involved in many inflammatory diseases; however, most CCR2 and CCR5 clinical candidates have been unsuccessful. (Pre)clinical evidence suggests that dual CCR2/CCR5 inhibition might be more effective in the treatment of such multifactorial diseases. In this regard, the highly conserved intracellular binding site in chemokine receptors provides a new avenue for the design of multitarget ligands. In this study, we synthesized and evaluated the biological activity of a series of triazolopyrimidinone derivatives in CCR2 and CCR5. Radioligand binding assays first showed that they bind to the intracellular site of CCR2, and in combination with functional assays on CCR5, we explored structure–affinity/activity relationships in both receptors. Although most compounds were CCR2-selective, 39 and 43 inhibited β-arrestin recruitment in CCR5 with high potency. Moreover, these compounds displayed an insurmountable mechanism of inhibition in both receptors, which holds promise for improved efficacy in inflammatory diseases.

Monte Carlo simulations are used to calculate the relative biological effectiveness (RBE) of 300 MeV u(-1) carbon-ion beams at different depths in a cylindrical water phantom of 10 cm radius and 30 cm long. RBE values for the induction of DNA double strand breaks (DSB), a biological endpoint closely related to cell inactivation, are estimated for monoenergetic and energy-modulated carbon ion beams. Individual contributions to the RBE from primary ions and secondary nuclear fragments are simulated separately. These simulations are based on a multi-scale modelling approach by first applying the FLUKA (version 2011.2.17) transport code to estimate the absorbed doses and fluence energy spectra, then using the MCDS (version 3.10A) damage code for DSB yields. The approach is efficient since it separates the non-stochastic dosimetry problem from the stochastic DNA damage problem. The MCDS code predicts the major trends of the DSB yields from detailed track structure simulations. It is found that, as depth is increasing, RBE values increase slowly from the entrance depth to the plateau region and change substantially in the Bragg peak region. RBE values reach their maxima at the distal edge of the Bragg peak. Beyond this edge, contributions to RBE are entirely from nuclear fragments. Maximum RBE values at the distal edges of the Bragg peak and the spread-out Bragg peak are, respectively, 3.0 and 2.8. The present approach has the flexibility to weight RBE contributions from different DSB classes, i.e. DSB0, DSB+ and DSB++.