A particle track-repeating algorithm for proton beam dose calculation

A particle track-repeating algorithm has been developed for proton beam dose calculation for radiotherapy. Monoenergetic protons with 250 MeV kinetic energy were simulated in an infinite water phantom using the GEANT3 Monte Carlo code. The changes in location, angle and energy for every transport step and the energy deposition along the track were recorded for the primary protons and all secondary particles. When calculating dose for a patient with a realistic proton beam, the pre-generated particle tracks were repeated in the patient geometry consisting of air, soft tissue and bone. The medium and density for each dose scoring voxel in the patient geometry were derived from patient CT data. The starting point, at which a proton track was repeated, was determined according to the incident proton energy. Thus, any protons with kinetic energy less than 250 MeV can be simulated. Based on the direction of the incident proton, the tracks were first rotated and for the subsequent steps, the scattering angles were simply repeated for air and soft tissue but adjusted properly based on the scattering power for bone. The particle step lengths were adjusted based on the density for air and soft tissue and also on the stopping powers for bone while keeping the energy deposition unchanged in each step. The difference in nuclear interactions and secondary particle generation between water and these materials was ignored. The algorithm has been validated by comparing the dose distributions in uniform water and layered heterogeneous phantoms with those calculated using the GEANT3 code for 120, 150, 180 and 250 MeV proton beams. The differences between them were within 2%. The new algorithm was about 13 times faster than the GEANT3 Monte Carlo code for a uniform phantom geometry and over 700 times faster for a heterogeneous phantom geometry.     

Decomposition of the telomere-targeting agent BRACO19 in physiological media results in products with decreased inhibitory potential

The stability of the acridine-based telomere-targeting agent BRACO19, a G-quadruplex stabilizing substance, was tested at different pH, temperature and in different dissolution media. Analysis was performed by HPLC. Decomposition products were examined by LC/MS and NMR. The TRAP assay was used to determine the inhibitory potential of the decomposition products on telomerase activity. The results show that the stability of BRACO19 strongly depends on pH and temperature. Decomposition was fastest at physiological pH and temperature while the type of dissolution medium had no major influence on stability. The most probable mechanism for this decomposition seems to be a hydrolysis of the amide bonds in position 3 and 6 of the acridine ring and/or a deamination of the phenyl ring. The decomposition products showed a reduced inhibitory potential compared to the parent compound BRACO19. The results demonstrate that the preparation of dosage forms and their storage conditions will have an important influence on the stability – and hence biological efficacy – of BRACO19 and related substances.     

Modulation of osteogenic potential by recombinant human bone morphogenic protein-2 in human periodontal ligament cells: effect of serum, culture medium, and osteoinductive medium

BACKGROUND AND OBJECTIVE: Bone morphogenic proteins are known, in animal models, to promote many developmental processes, including osteogenesis. Clinical trials are currently underway to evaluate the potential of bone morphogenic proteins to promote bone and periodontal regeneration in humans. The aim of this study was to establish an optimal cell culture condition for using to study the biological effects of recombinant human bone morphogenic protein-2 on periodontal ligament cells. MATERIAL AND METHODS: The roles of serum concentration, types of culture medium (alpha-modified essential medium or Dulbecco's modified Eagle's medium), the presence of osteoinductive medium (including dexamethasone, ascorbic acid and beta-glycerophosphate), and timing of addition of the osteoinductive medium and recombinant human bone morphogenic protein-2, on the expression of alkaline phosphatase were investigated in cultured periodontal ligament cells. Cytochemical stainings and biological assay of alkaline phosphatase were also demonstrated. RESULTS: Our results suggested that an increased concentration of serum might mask the effect of recombinant human bone morphogenic protein-2 on the expression of alkaline phosphatase in periodontal ligament cells. alpha-Modified essential medium was found to induce a stronger cytochemical staining of the alkaline phosphatase than Dulbecco's modified Eagle's medium under similar culture conditions. Pre-incubation of cells with osteoinductive medium before the addition of various concentrations of recombinant human bone morphogenic protein-2 enhanced greater alkaline phosphatase expression than the simultaneous presence of both osteoinductive medium and recombinant human bone morphogenic protein-2. CONCLUSION: The findings of this study suggest that the effect of recombinant human bone morphogenic protein-2 on periodontal ligament cells could be efficiently investigated after the proper selection of culture variables and temporal sequence of adding bioactive factors. The optimal culture condition identified in this study might be useful in further studies to elucidate the regulatory mechanism of periodontal ligament cells in periodontal regeneration after stimulation with recombinant human bone morphogenic protein-2.     

Functional characterisation of a complex mutation in the α(1,4)galactosyltransferase gene in Taiwanese individuals with p phenotype

Background and objective: Individuals with p phenotype lack P1, P^k and P antigens on red blood cells, presumably as a result of deficiency in the enzyme α(1,4)galactosyltransferase (A4GALT). The aim of this study was to explore the molecular background of a Taiwanese family with p phenotype. Materials and methods: Blood samples from two p siblings and seven family members were investigated. The coding region of the A4GALT gene was analysed by polymerase chain reaction and direct sequencing. The wild‐ and mutant‐complementary DNAs (cDNAs) of A4GALT were cloned into an expression vector and transfected to Chinese hamster ovary (CHO) cells. P^k expression on the transfected cells was analysed by flow cytometry and the activities of A4GALT were measured by high‐performance liquid chromatography. Results: The two individuals with p phenotype were homozygous for the complex mutation, which was caused by a combined deletion and insertion between nt 418 and 428. No expression of P^k and no enzyme activity were observed in cells transfected with the mutant construct. Conclusion: The first case of p phenotype in Taiwan was caused by a non‐functional allele resulting from a homozygous complex mutation of A4GALT gene.     

Ultra-thin TLDs for skin dose determination in high energy photon beams

Estimation of surface dose is very important for patients undergoing radiation therapy. In this work we investigate the dose at the surface of a water phantom and at a depth of 0.007 cm, the practical reference depth for skin as recommended by ICRP and ICRU, with ultra-thin TLDs and Monte Carlo calculations. The calculations and measurements were carried out for fields ranging from 5 x 5 cm2 to 20 x 20 cm2 for 6 MV, 10 MV and 18 MV photon beams. The variation of the surface dose with angle of incidence and field size was investigated. Also, the exit dose was computed and measured for the same fields and angles of incidence. The dose at the ICRU reference depth was computed. Good agreement (+/-5%) was achieved between measurements and calculations. The surface dose at the entrance increased with the angle of incidence and/or the field size. The exit dose decreased with the angle of incidence but it increased with field size. The dose at the surface of the patient is mostly dependent on the beam energy, modality and beam obliquity rather than the field size and field separation. By correlating TLD measurements with Monte Carlo calculations, we were able to predict the dose at the skin surface with good accuracy. Knowing the dose received at the surface of the patient can lead to prediction of skin reactions helping with the design of new treatment techniques and alternative dose fractionation schemes.     

The HPV E6 oncoprotein targets histone methyltransferases for modulating specific gene transcription

Expression of viral proteins causes important epigenetic changes leading to abnormal cell growth. Whether viral proteins directly target histone methyltransferases (HMTs), a key family enzyme for epigenetic regulation, and modulate their enzymatic activities remains elusive. Here we show that the E6 proteins of both low-risk and high-risk human papillomavirus (HPV) interact with three coactivator HMTs, CARM1, PRMT1 and SET7, and downregulate their enzymatic activities in vitro and in HPV-transformed HeLa cells. Furthermore, these three HMTs are required for E6 to attenuate p53 transactivation function. Mechanistically, E6 hampers CARM1- and PRMT1-catalyzed histone methylation at p53-responsive promoters, and suppresses the binding of p53 to chromatinized DNA independently of E6-mediated p53 degradation. p53 pre-methylated at lysine-372 (p53K372 mono-methylation) by SET7 protects p53 from E6-induced degradation. Consistently, E6 downregulates p53K372 mono-methylation and thus reduces p53 protein stability. As a result of the E6-mediated inhibition of HMT activity, expression of p53 downstream genes is suppressed. Together, our results not only reveal a clever approach for the virus to interfere with p53 function, but also demonstrate the modulation of HMT activity as a novel mechanism of epigenetic regulation by a viral oncoprotein.     

Technical quality of root canal treatment in Taiwan

AIM: To evaluate the current technical quality of root canal treatment (RCT) in Taiwan. METHODOLOGY: A total of 1085 RCT cases, randomly selected from a large sample and representative of the Taiwanese population from April to September 2000, were evaluated by eight endodontic specialists. The qualitative evaluation of RCT cases was based on two variables: length of the root filling and density of the obturation. A root canal with both adequate filling length (the apical termination of the root filling within 2 mm of the radiographic apex) and complete obturation (no lateral or apical canal lumen visible in the apical one-third of the root canal) was defined as having good-quality endodontic work (GQEW). A tooth was defined as having a GQEW when all its canals were categorized as GQEW. RESULTS: From a total of 1867 root canals, overfilling occurred in 235 (12.6%), adequate filling length in 1152 (61.7%), underfilling in 466 (25.0%) and no filling in 12 (0.6%). Of the 1867 root canals, 710 (38.0%) demonstrated complete obturation and 1157 (62%) demonstrated incomplete obturation. GQEW was found in 650 (34.8%) root canals and 329 (30.3%) teeth. The percentage of teeth with GQEW in hospital cases (38.1%) was significantly greater (P < 0.001) than that in private clinic cases (24.3%). In addition, the frequency of teeth with GQEW in the anterior teeth (40.4%) or in the premolars (33%) was significantly greater (P < 0.001) than that in the molars (18.4%). CONCLUSIONS: Approximately 70% of the teeth receiving RCT in Taiwan were either of inadequate filling length or sealing density.