Cytotoxic 2-benzylidene-6-(nitrobenzylidene)cyclohexanones which display substantially greater toxicity for neoplasms than non-malignant cells

Various 2-benzylidene-6-(nitrobenzylidene)cyclohexanones were prepared as candidate cytotoxins in which the nitro group was located in the ortho, meta and para positions leading to series 1–3, respectively. The CC₅₀ values towards human HSC-2 and HSC-4 oral squamous cell carcinomas as well as human HL-60 promyelocytic leukemic cells are in the low micromolar range in general. On the other hand, most of the compounds afforded clear evidence of being far less toxic towards human HGF gingival fibroblasts, HPC pulp cells and HPLF periodontal ligament fibroblasts which are non-malignant cells. Selectivity index (SI) figures were generated which are the ratios of the average CC₅₀ values towards normal cells and the CC₅₀ figure towards a malignant cell line. Huge SI values were obtained for many of the compounds. In particular 1c, 2f, 3c and 3g which have average SI values of >76, >38, 124 and 341, respectively, are clearly lead molecules affording direction for amplification of this area of study. A lead compound 1c caused internucleosomal DNA fragmentation and activation of caspase-3 in HL-60 cells but not in HSC-2 carcinomas. In a short-term toxicity study, doses up to and including 300 mg/kg of the majority of the compounds prepared in this study did not cause any mortalities to mice. Some guidelines for development of these tumor-selective cytotoxins are presented.     

Monte Carlo evaluation of occupational exposure during uterine artery embolization

PurposeUterine fibroids affect women mainly of childbearing age, an alternative for the treatment of these fibroids is uterine artery embolization (UAE), a minimally invasive procedure which uses fluoroscopy, providing radiation doses often high, due to the fact that professionals remain in the room throughout the procedure. In this work, equivalent and effective doses were evaluated for the main physician, for the assistant and for the patient during the UAE procedure.MethodsDoses were calculated using computer simulation with the Monte Carlo Method, and virtual anthropomorphic phantoms, in a typical scenario of interventional radiology with field sizes of 20 × 20, 25 × 25 and 32 × 32 cm², tube voltages of 70, 80, 90 and 100 kV, and projections of LAO45, RAO45 and PA.ResultsThe results showed that the highest doses received by the professionals were for the LAO45 projection with 32 × 32 cm² field size and 100 kV tube voltage, which is in accordance with the existing literature. The highest equivalent doses, without the protective equipment, were in the eyes, skin, breast and stomach for the main physician, and for the assistant they were in the eyes, breast, thyroid and skin. When she used the protective equipment, the highest equivalent doses for the main physician were on the skin, brain, bone marrow and bone surface, and for the assistant they were on the skin, brain, red bone marrow and bone surface.ConclusionsEffective doses increased up to 3186% for the main physician, and 2462% for the assistant, without protective equipment, thus showing their importance.     

New Approach to Develop Optimized Sunscreens that Enable Cutaneous Vitamin D Formation with Minimal Erythema Risk

Sunscreens protect the skin against erythemal radiation (E_er). But at the same time they reduce the effective radiation dose (E_VD) responsible for the formation of previtamin D in the skin. The paper describes a calculation method for optimizing the ratio E_VD/E_er behind sunscreens e.g. with SPF 5, 15 and 30 respectively. Taking into account that a majority of people in industrialized countries suffer from a shortage in vitamin D even in summer time, the ratio E_vd/E_er is a new and important criterion for the quality of sunscreens. Furthermore the exposure time t_vd needed per day for forming the equivalent of the recommended amount of 2000 IU of vitamin D per day for skin type 2 is estimated when sunscreens with different filter compositions are used. In vitro experiments show a significant increase of the conversion of 7-dehydrocholesterol (7-DHC) to previtamin D when exposed to artificial solar radiation behind an experimental sunscreen optimized for previtamin D production compared to a commercial sunscreen having the same SPF.     

Chemical composition and enzymic activity of some worm casts

Summary An investigation was carried out to analyze the chemical composition and enzymatic activity of five worm casts obtained from various animal dungs and municipal waste. The E₄/E₆ values for humic and fulvic acids were higher in all worm casts than in a typical Umbrian soil. Comparisons between worm cast, substrate and soil IR and NMR spectra demonstrated the humifying capacity of worms. Enzyme activity was high in all casts and only dehydrogenase seemed to be affected by lead concentration.Research work supported by CNR, Italy. Special grant I.P.R.A.-Sub-project 1. Paper N. 94.     

Radiation dose in repeated CT guided radiofrequency ablations

ObjectiveTo calculate the cumulative effective and skin doses in patients that underwent repeated CT guided radiofrequency ablations (RFA).Materials and methodsFrom all patients that had undergone RFA during a five years period those which had three or more RFAs were selected. Using the CT images DICOM data, the dose length product (DLP), effective dose (E), skin dose profiles as well as the peak skin dose (PSD) were calculated, using appropriate methods and software developed for this purpose. For each patient, cumulative DLP and E were also calculated from the sum of the respective figures of each individual procedure. To calculate PSD, the skin dose profiles of each procedure were overlaid on the same Z-axis scale using anatomical landmarks for reference and the skin doses to each point were summed up.ResultsFive patients were studied; four had undergone 3 RFAs and one 10 RFAs. Cumulative DLP, E and PSD ranges were 5.6–22.3 Gy cm, 0.08–0.36 Sv and 0.8–3.4 Gy, respectively. Median E and PSD values per RFA were 35 mSv and 0.4 Gy, respectively. For comparison purposes it must be noted that in this CT department a routine abdomen-pelvis scan results to an E of about 10 mSv.ConclusionsPatients that undergo repeated RFAs are exposed to considerably high radiation exposure levels. When these patients are in the final stage of malignant diseases, stochastic effects may not be of major concern. However, optimization of the exposure factors and monitoring of these patients to avoid skin injuries are required.     

A New Approach to Reduce Uncertainties in Space Radiation Cancer Risk Predictions

The prediction of space radiation induced cancer risk carries large uncertainties with two of the largest uncertainties being radiation quality and dose-rate effects. In risk models the ratio of the quality factor (QF) to the dose and dose-rate reduction effectiveness factor (DDREF) parameter is used to scale organ doses for cosmic ray proton and high charge and energy (HZE) particles to a hazard rate for γ-rays derived from human epidemiology data. In previous work, particle track structure concepts were used to formulate a space radiation QF function that is dependent on particle charge number Z, and kinetic energy per atomic mass unit, E. QF uncertainties where represented by subjective probability distribution functions (PDF) for the three QF parameters that described its maximum value and shape parameters for Z and E dependences. Here I report on an analysis of a maximum QF parameter and its uncertainty using mouse tumor induction data. Because experimental data for risks at low doses of γ-rays are highly uncertain which impacts estimates of maximum values of relative biological effectiveness (RBE_max), I developed an alternate QF model, denoted QF_γAcute where QFs are defined relative to higher acute γ-ray doses (0.5 to 3 Gy). The alternate model reduces the dependence of risk projections on the DDREF, however a DDREF is still needed for risk estimates for high-energy protons and other primary or secondary sparsely ionizing space radiation components. Risk projections (upper confidence levels (CL)) for space missions show a reduction of about 40% (CL∼50%) using the QF_γAcute model compared the QFs based on RBE_max and about 25% (CL∼35%) compared to previous estimates. In addition, I discuss how a possible qualitative difference leading to increased tumor lethality for HZE particles compared to low LET radiation and background tumors remains a large uncertainty in risk estimates.     

Altered kinetic properties of liver mitochondrial membrane-bound enzyme activities following paracetamol hepatotoxicity in the rat

The effects of treatment with subtoxic (375 mg/kg) and toxic (750 mg/kg) doses of paracetamol on NADH oxidase, succinoxidase and Mg²⁺-ATPase activities in rat liver submitochondrial particles were examined. In the NADH oxidase system, treatment with subtoxic doses of paracetamol resulted in a 37% increase in activation energy in the high temperature range (E¹) while the phase transition temperature (Tt) for this system decreased by 9‡C. Subtoxic doses caused a 43% decrease in E₁. For the succinoxidase system, Tt decreased by 2.4 to 3.4‡C after paracetamol administration. E₂ increased by 42% only in the subtoxic-treatment group while E₁ remained unaltered in both paracetamol-treated groups. For the Mg²⁺-ATPase system, subtoxic doses of paracetamol treatment did not change the values of E₁ E₂ and Tt whereas toxic dose treatment resulted in a 29% decrease in E₂ with a concomitant increase in Tt by 2.4‡C without any change in the value of E₁ The results thus suggest that treatment with toxic and subtoxic doses of paracetamol results in possible differential alterations in the membrane lipid milieu.     

Effective dose in percutaneous transhepatic biliary drainage examination using PCXMC2.0 and MCNP5 Monte Carlo codes

ObjectivesTo estimate the organ equivalent doses and the effective doses (E) in patient undergoing percutaneous transhepatic biliary drainage (PTBD) examinations, using the MCNP5 and PCXMC2 Monte Carlo-based codes.MethodsThe purpose of this study is to estimate the organ doses to patients undergoing PTBD examinations by clinical measurements and Monte Carlo simulation. Dose area products (DAP) values were assessed during examination of 43 patients undergoing PTBD examination separated into groups based on the gender and the dimensions and location of the beam.ResultsMonte Carlo simulation of photon transport in male and female mathematical phantoms was applied using the MCNP5 and PCXMC2 codes in order to estimate equivalent organ doses. Regarding the PTBD examination the organ receiving the maximum radiation dose was the lumbar spine. The mean calculated H_T for the lumbar spine using the MCNP5 and PCXMC2 methods respectively, was 117.25 mSv and 131.7 mSv, in males. The corresponding doses were 139.45 mSv and 157.1 mSv respectively in females. The H_T values for organs receiving considerable amounts of radiation during PTBD examinations were varied between 0.16% and 73.2% for the male group and between 1.10% and 77.6% for the female group. E in females and males using MCNP5 and PCXMC2.0 was 5.88 mSv and 6.77 mSv, and 4.93 mSv and 5.60 mSv.ConclusionThe doses remain high compared to other invasive operations in interventional radiology. There is a reasonable good coincidence between the MCNP5 and PCXMC2.0 calculation for most of the organs.     

Role of differential adhesion in cell cluster evolution: from vasculogenesis to cancer metastasis

Cell–cell and cell–matrix adhesions are fundamental to numerous physiological processes, including angiogenesis, tumourigenesis, metastatic spreading and wound healing. We use cellular potts model to computationally predict the organisation of cells within a 3D matrix. The energy potentials regulating cell–cell (J_CC) and cell–matrix (J_MC) adhesive interactions are systematically varied to represent different, biologically relevant adhesive conditions. Chemotactically induced cell migration is also addressed. Starting from a cluster of cells, variations in relative cell adhesion alone lead to different cellular patterns such as spreading of metastatic tumours and angiogenesis. The combination of low cell–cell adhesion (high J_CC) and high heterotypic adhesion (low J_MC) favours the fragmentation of the original cluster into multiple, smaller cell clusters (metastasis). Conversely, cellular systems exhibiting high-homotypic affinity (low J_CC) preserve their original configuration, avoiding fragmentation (organogenesis). For intermediate values of J_CC and J_MC (i.e. J_CC/J_MC ～ 1), tubular and corrugated structures form. Fully developed vascular trees are assembled only in systems in which contact-inhibited chemotaxis is activated upon cell contact. Also, the rate of secretion, diffusion and sequestration of chemotactic factors, cell deformability and motility do not significantly affect these trends. Further developments of this computational model will predict the efficacy of therapeutic interventions to modulate the diseased microenvironment by directly altering cell cohesion.     

A Process Similar to Autophagy Is Associated with Cytocidal Chloroquine Resistance in Plasmodium falciparum

Resistance to the cytostatic activity of the antimalarial drug chloroquine (CQ) is becoming well understood, however, resistance to cytocidal effects of CQ is largely unexplored. We find that PfCRT mutations that almost fully recapitulate P. falciparum cytostatic CQ resistance (CQR^CS) as quantified by CQ IC₅₀ shift, account for only 10–20% of cytocidal CQR (CQR^CC) as quantified by CQ LD₅₀ shift. Quantitative trait loci (QTL) analysis of the progeny of a chloroquine sensitive (CQS; strain HB3)×chloroquine resistant (CQR; strain Dd2) genetic cross identifies distinct genetic architectures for CQR^CS vs CQR^CC phenotypes, including identification of novel interacting chromosomal loci that influence CQ LD₅₀. Candidate genes in these loci are consistent with a role for autophagy in CQR^CC, leading us to directly examine the autophagy pathway in intraerythrocytic CQR parasites. Indirect immunofluorescence of RBC infected with synchronized CQS vs CQR trophozoite stage parasites reveals differences in the distribution of the autophagy marker protein PfATG8 coinciding with CQR^CC. Taken together, the data show that an unusual autophagy – like process is either activated or inhibited for intraerythrocytic trophozoite parasites at LD₅₀ doses (but not IC₅₀ doses) of CQ, that the pathway is altered in CQR P. falciparum, and that it may contribute along with mutations in PfCRT to confer the CQR^CC phenotype.     

Patient doses and occupational exposure in a hybrid operating room

PurposeThis study aimed to characterize the radiation exposure to patients and workers in a new vascular hybrid operating room during X-ray-guided procedures.MethodsDuring one year, data from 260 interventions performed in a hybrid operating room equipped with a Siemens Artis Zeego angiography system were monitored. The patient doses were analysed using the following parameters: radiation time, kerma-area product, patient entrance reference point dose and peak skin dose. Staff radiation exposure and ambient dose equivalent were also measured using direct reading dosimeters and thermoluminescent dosimeters.ResultsThe radiation time, kerma-area product, patient entrance reference point dose and peak skin dose were, on average, 19:15 min, 67 Gy·cm², 0.41 Gy and 0.23 Gy, respectively. Although the contribution of the acquisition mode was smaller than 5% in terms of the radiation time, this mode accounted for more than 60% of the effective dose per patient. All of the worker dose measurements remained below the limits established by law.ConclusionsThe working conditions in the hybrid operating room HOR are safe in terms of patient and staff radiation protection. Nevertheless, doses are highly dependent on the workload; thus, further research is necessary to evaluate any possible radiological deviation of the daily working conditions in the HOR.     

Synthesis of Camphecene and Cytisine Conjugates Using Click Chemistry Methodology and Study of Their Antiviral Activity

A series of camphecene and quinolizidine alkaloid (?)‐cytisine conjugates has been obtained for the first time using ‘click’ chemistry methodology. The cytotoxicity and virus‐inhibiting activity of compounds were determined against MDCK cells and influenza virus A/Puerto Rico/8/34 (H1N1), correspondingly, in in vitro tests. Based on the results obtained, values of 50 % cytotoxic dose (CC₅₀), 50 % inhibition dose (IC₅₀) and selectivity index (SI) were determined for each compound. It has been shown that the antiviral activity is affected by the length and nature of linkers between cytisine and camphor units. Conjugate 13 ((1R,5S)‐3‐(6‐{4‐[(2‐{(E)‐[(1R,4R)‐1,7,7‐trimethylbicyclo[2.2.1]heptan‐2‐ylidene]amino}ethoxy)methyl]‐1H‐1,2,3‐triazol‐1‐yl}hexyl)‐1,2,3,4,5,6‐hexahydro‐8H‐1,5‐methanopyrido[1,2‐a][1,5]diazocin‐8‐one), which contains cytisine fragment separated from triazole ring by –C₆H₁₂– aliphatic linker, showed the highest activity at relatively low toxicity (CC₅₀=168 μmol, IC₅₀=8 μmol, SI=20). Its selectivity index appeared higher than that of reference compound, rimantadine. According to theoretical calculations, the antiviral activity of the lead compound 13 can be explained by its influence on the functioning of neuraminidase.     

On the relationship between cumulative correlation coefficients and the quality of crystallographic data sets

In 2012, Karplus and Diederichs demonstrated that the Pearson correlation coefficient CC_1/2 is a far better indicator of the quality and resolution of crystallographic data sets than more traditional measures like merging R‐factor or signal‐to‐noise ratio. More specifically, they proposed that CC_1/2 be computed for data sets in thin shells of increasing resolution so that the resolution dependence of that quantity can be examined. Recently, however, the CC_1/2 values of entire data sets, i.e., cumulative correlation coefficients, have been used as a measure of data quality. Here, we show that the difference in cumulative CC_1/2 value between a data set that has been accurately measured and a data set that has not is likely to be small. Furthermore, structures obtained by molecular replacement from poorly measured data sets are likely to suffer from extreme model bias.     

Chromosome aberration analysis and the influence of mitotic delay after simulated partial-body exposure with high doses of sparsely and densely ionising radiation

The influence of high doses of sparsely and densely ionising radiation on the yield of aberrant human peripheral lymphocytes in simulated partial-body exposures was studied by investigating radiation-induced chromosome aberration frequencies, namely dicentric and centric ring chromosomes. Peripheral blood samples from two volunteers were irradiated with high doses of 200 kV X-rays or neutrons with a mean energy of <E _n>=2.1 MeV and partial-body exposure was simulated by mixing irradiated and non-irradiated blood from the same two donors in proportions of 25, 50, and 75%. Lymphocytes were cultured and first-division metaphase cells were collected after culture times of 48, 56, and 72 h. A significant underrepresentation of dicentric and centric ring chromosomes was observed at the three highest doses of X-rays between the different culture times for nearly all proportions. After neutron irradiation, some significant differences were observed at all doses and all culture times, without however, revealing any systematic pattern. The distribution of dicentric and ring chromosomes showed overdispersion for both radiation types. After simulated partial-body exposures with 200 kV X-rays and <E _n>=2.1 MeV neutrons, strong mitotic delays could be observed, which depended on both the irradiated volume and the applied dose: the smaller the irradiated volume and the higher the dose, the higher was the selective advantage of non-irradiated cells. For the purpose of biological dosimetry after partial body exposure, an extension of the lymphocyte culture time is suggested at least for doses ≥3.0 Gy of 200 kV X-rays and ≥0.5 Gy of <E _n>=2.1 MeV neutrons in order to prevent a systematic underestimation of cytogenetic damage.     

Quantitative Lesion-to-Fat Elasticity Ratio Measured by Shear-Wave Elastography for Breast Mass: Which Area Should Be Selected as the Fat Reference?

Objectives

To investigate whether the diagnostic performance of lesion-to-fat elasticity ratio (E_ratio) was affected by the location of the reference fat.

Methods

For 257 breast masses in 250 women who underwent shear-wave elastography before biopsy or surgery, multiple E_ratios were measured with a fixed region-of-interest (ROI) in the mass along with multiple ROIs over the surrounding fat in different locations. Logistic regression analysis was used to determine that E_ratio was independently associated with malignancy adjusted for the location of fat ROI (depth, laterality, and distance from lesion or skin). Mean (E_mean) and maximum (E_max) elasticity values of fat were divided into four groups according to their interquartile ranges. Diagnostic performance of each group was evaluated using the area under the ROC curve (AUC). False diagnoses of E_ratio were reviewed for ROIs on areas showing artifactual high or low stiffness and analyzed by logistic regression analysis to determine variables (associated palpable abnormality, lesion size, the vertical distance from fat ROI to skin, and elasticity values of lesion or fat) independently associated with false results.

Results

E_ratio was independently associated with malignancy adjusted for the location of fat ROI (P<0.0001). Among four groups of fat elasticity values, the AUC showed no significant difference (<25th percentile, 25th percentile~median, median~75th percentile, and ≥75th percentile; 0.973, 0.982, 0.967, and 0.954 for E_mean; 0.977, 0.967, 0.966, and 0.957 for E_max). Fat elasticity values were independently associated with false results of E_ratio with the cut-off of 3.18 from ROC curve (P<0.0001). ROIs were set on fat showing artifactual high stiffness in 90% of 10 false negatives and on lesion showing vertical striped artifact or fat showing artifactual low stiffness in 77.5% of 71 false positives.

Conclusion

E_ratio shows good diagnostic performance regardless of the location of reference fat, except when it is placed in areas of artifacts.     

1-(3-Aminomethyl-4-hydroxyphenyl)-3-pyridinyl-2-propen-1-ones: A novel group of tumour-selective cytotoxins

Two series of 1-(3-aminomethyl-4-hydroxyphenyl)-3-pyridinyl-2-propen-1-ones, designed as novel cytotoxins, were synthesized. The compounds had low CC₅₀ values in the micromolar range against HL-60 promyelocytic leukemic cells and HSC-2, HSC-3 and HSC-4 oral squamous cell carcinomas. The CC₅₀ values of these compounds were higher towards non-malignant HGF (gingival fibroblasts), HPC (pulp cells), and HPLF (periodontal ligament fibroblasts) cells, which reveals the tumour-selectivity of these enones. A representative compound 4c caused cleavage of PARP1 in HSC-2 cells but not in HGF cells, which may be a contributing factor to the tumour-selectivity.     

Esterase D polymorphism in a French-Canadian population

Summary Red blood cell esterase D (ESD) polymorphism was studied in a French-Canadian population from Quebec city, Canada, by means of high voltage electrophoresis on agarose gel followed, in heterozygotes for ESD1, by IEF to reveal the possible allele ESD^*5. Frequencies of the ESD alleles in 904 unrelated individuals were ESD^*1: 0.888, EDS^*2: 0.095 and ESD^*5: 0.017. The segregation pattern observed in 275 families confirmed a Mendelian inheritance of three autosomal alleles.     

Tumour-specific cytotoxicity and structure–activity relationships of novel 1-[3-(2-methoxyethylthio)propionyl]-3,5-bis(benzylidene)-4-piperidones

A series of 1-acyl-3,5-bis(benzylidene)-4-piperidones 3–7 were designed and synthesized as novel cytotoxic agents. These compounds displayed potent cytotoxic properties towards human Molt4/C8, CEM, HSC-2, HSC-3 and HSC-4 neoplasms and also to murine L1210 cells. The majority of the compounds have sub-micromolar or very low micromolar IC₅₀ and CC₅₀ values and are significantly more potent than the reference alkylating drug melphalan. Evaluation of these compounds against non-malignant HGF and HPLF cells revealed the tumour-specific toxicity. In particular, 3e emerged as a promising lead cytotoxic agent which caused apoptosis and PARP1 cleavage in HSC-2 cells.     

Measuring skin dose in CT examinations under complex geometries: Instruments,methods and considerations

ObjectiveTo investigate skin dose in Computed Tomography (CT) and its dependence on scanning geometry.Materials and methodsMeasurements of entrance surface air kerma (ESAK) in free air and entrance skin dose (ESD) on an anthropomorphic phantom were performed in a 64-slice CT scanner, using two different instruments: the Dose Profiler (DP) and the QED skin diode (QEDSD). Using DP and QEDSD, the ESAK rate profiles at the isocenter and at different distances from it, were measured using axial scans. Using DP and helical scans the ESAK rate profile in the Z-axis was acquired. The same profile was acquired with the QEDSD also, using many axial scans and manual table translation. ESD measurements were performed with the DP and QEDSD, in axial and helical scan mode.ResultsESAK measurements with DP and QEDSD were in good agreement, for both point dose and profile measurements. The agreement was also good for ESD measurements but not for helical scans, due to variable X-ray beam overlapping and different tube angular positions at each scan start. It was observed that the ESD values at different Y-axis offsets were comparable to the respective ESAK values recorded at the same Y-axis offset distances without the phantom.ConclusionsBoth DP and QEDSD were proven suitable for performing point ESD measurements. However, calculating the skin dose distribution in CT examinations is a very challenging task. A practical approach would be for CT scanners to provide a conservative estimate of the peak skin dose using the isocenter ESAK value.