Measurements of the morphology of the ankle joint, performed mostly for surgical planning of total ankle arthroplasty and for collecting data for total ankle prosthesis design, are often made on planar radiographs, and therefore can be very sensitive to the positioning of the joint during imaging. The current study aimed to compare ankle morphological measurements using CT-generated 2D images with gold standard values obtained from 3D CT data; to determine the sensitivity of the 2D measurements to mal-positioning of the ankle during imaging; and to quantify the repeatability of the 2D measurements under simulated positioning conditions involving random errors.Fifty-eight cadaveric ankles fixed in the neutral joint position (standard pose) were CT scanned, and the data were used to simulate lateral and frontal radiographs under various positioning conditions using digitally reconstructed radiographs (DRR).In the standard pose for imaging, most ankle morphometric parameters measured using 2D images were highly correlated (R > 0.8) to the gold standard values defined by the 3D CT data. For measurements made on the lateral views, the only parameters sensitive to rotational pose errors were longitudinal distances between the most anterior and the most posterior points of the tibial mortise and the tibial profile, which have important implications for determining the optimal cutting level of the bone during arthroplasty. Measurements of the trochlea tali width on the frontal views underestimated the standard values by up to 31.2%, with only a moderate reliability, suggesting that pre-surgical evaluations based on the trochlea tali width should be made with caution in order to avoid inappropriate selection of prosthesis sizes.While highly correlated with 3D morphological measurements, some 2D measurements were affected by the bone poses in space during imaging, which may affect surgical decision-making in total ankle arthroplasty, including the amount of bone resection and the selection of the implant sizes. The linear regression equations for the relationship between 2D and 3D measurements will be helpful for correcting the errors in 2D morphometric measurements for clinical applications. Total ankle arthroplasty (TAA) is an important treatment option for the management of advanced ankle osteoarthritis, a disease often leading to impairment of locomotion, physical disability, and reduced quality of life ,2. CliniIt is well recognized that conformity of a TAA design to the bone morphology, including the proper sizing of the components, is an important factor for the success of the prosthesis in replicating the function of the joint . For exaAmong the clinically available medical imaging modalities, bi-planar radiographs are commonly used for this purpose owing to their convenience, low cost, and low radiation dose compared with other modalities such as MRI or CT . Since bFor a morphological measurement of the ankle on a radiograph to be accurate, the true positions of anatomical bony landmarks should be identifiable on the radiograph, and the bones have to be positioned correctly with respect to the X-ray source to avoid projecting oblique shapes and overlapping surfaces onto the radiograph, i.e., radiographs of the ankle must be A/P, M/L or M views. However, this ideal positioning is difficult to achieve in clinical practice. The positioning error of the ankle as a result of contact of the exterior shape of the lower limb with the image plane, or improper position of the beam focus by an operator during the radiography procedure might cause artificial projection errors on the planar radiograph ,19.Computerized Tomography (CT) scans can be used to obtain more accurate morphological data of the ankle bones  than thoThe purposes of this study were (a) to compare the measurements of ankle morphology in the standard pose using 3D CT images with those using 2D images, and to determine the relationships between 2D and 3D measurements using linear regression analysis; (b) to determine the sensitivity of the 2D measurements to the errors associated with the mal-positioning of the ankle during imaging; (c) to quantify the repeatability of the 2D measurements under simulated positioning conditions involving random errors; and (d) to evaluate the performance of the linear regression equations obtained in (a) in predicting 3D measurements from 2D measurements.The general procedural framework for the current study is shown in Figure , in a plastic frame .Fifty-eight fresh frozen ankle specimens, 22 females and 36 males, were used in the current study Table . These set al. [Two types of 3D models of the bones, i.e., volumetric and surface models, together with the plastic frame were reconstructed from the CT images following Lin et al. . The volet al. ,20 for tGiven the positions of the X-ray source and a CT-derived volumetric bone model in space with respective to the image plane, the DRR of the bone was generated by casting rays through the volume of the bone model . Each ofMeasurements of the morphological parameters on planar radiographs were first performed with the ankle model in the standard pose for imaging, with the anatomical axes parallel with those of the X-ray system depending on the direction of the imaging Figure . For theA systematic sensitivity analysis was performed to determine the changes in the 2D measurements as a result of small deviations about each of the three anatomical axes of the ankle specimen from the standard pose. From the standard pose, the model of the ankle specimen was then rotated by 6 degrees at 1-degree intervals in both directions about each of the three axes of the coordinate system, respectively Figure . The maxFor the repeatability of the 2D measurements, each specimen was perturbed from its standard pose with random errors in the three individual rotational components to obtain ten new poses. The random errors were generated within the above-mentioned maximum ranges. For each specimen 2D morphological parameters were measured from the DRRs of the ten different specimen model poses for each of the M/L and A/P views. Repeatability of the measurements of each parameter was then assessed in terms of the Intra-Class Correlation coefficients using a 2-way mixed-effects average model  for intra-examiner assessment . The valFor evaluating the performance of the linear regression equations derived from data in the standard pose for predicting the 3D measurements from 2D measurements, each specimen was perturbed from its standard pose with random errors in the three individual rotational components to obtain two sets of ten new poses, one set with random errors within a maximum of 3 degrees and another set within 6 degrees. The DRRs of these perturbed poses in the M/L and A/P views were used to obtain 2D measurements of the morphological parameters. These measurements were then corrected using the corresponding linear regression equations to give corrected 2D measurements. The residual errors of the corrected 2D measurements were calculated by comparing them to the 3D measurements. All the statistical analysis was performed using SPSS 13.0 .The results of the Shapiro-Wilk test showed that the parameters all had a normal distribution. In the standard pose without any positioning errors, most morphological parameters were significantly different between 2D and 3D measurements, except APA, MTiTh, MDV, TaR, MLATi and TaW  Table . For thoWith a rotational perturbation of 6 degrees in both directions about the Z-axis , the most sensitive parameter again was MLATa, overestimating the standard values by 145.3% for positive rotational perturbations and by 87.0% for negative rotational perturbations , TiSR (0.95), APG (0.97), APA (0.97), MTiTh (0.95), TaAL (0.92), TaR (0.95), TiW (0.97), MalW (0.91) and MLATa (0.89). Parameters with good reliability were MLATi (0.78), MDA (0.79) and MDV (0.65). The only parameter with a moderate reliability was TaW (ICC = 0.55).The linear regression equations derived from data in the standard pose performed quite well for poses of the specimen models that were perturbed within 3 degrees in the three individual rotational components. After correction with the linear regression equations Table . Pose erThe current study aimed to compare the measurements of ankle morphology in the standard pose using 3D CT images with those using 2D images, and to determine the relationships between 2D and 3D measurements using linear regression analysis; to determine the sensitivity of the 2D measurements to mal-positioning of the ankle during imaging; to quantify the repeatability of the 2D measurements under simulated positioning conditions involving random errors; and to evaluate the performance of the linear regression equations in predicting 3D measurements from 2D measurements. In the standard pose, six out of fourteen 2D morphological parameters were not significantly different from the 3D gold standard values, and most of the parameters were highly correlated between 2D and 3D measurements. Most 2D measurements were not sensitive to rotational perturbations, except for TaW, MLATi and MLATa to perturbations about the X-axis, and MDA, MDV and MLATa to perturbations about the Z-axis. Most of the 2D measurements were repeatable when subjected to randomized perturbations about each anatomical axis, except for TaW that had only moderate repeatability. The linear regression equations between the 2D and 3D measurements were shown to be effective in correcting the errors in the 2D morphometric measurements, which will be helpful for improved estimation of the morphometric parameters for clinical purposes.In the standard pose for imaging, most morphometric parameters measured using 2D images were highly correlated to the gold standard values defined by the 3D CT data. This was expected because in the standard imaging pose the morphometric parameters measured from the 2D images were related to the 3D data via the principle of perspective projection with only little variation among subjects. Nonetheless, significant differences between 2D and 3D measurements did exist in several morphometric parameters. On the other hand, for those few morphometric parameters, namely MTiTh, MDV and TaR, that had poor to moderate correlations with the 3D gold standard, no significant differences were found between their 2D and 3D measurements. This indicates that if the imaging is done in the standard pose, the 2D measurements can be regarded as good representations of the parameters as those measured using 3D CT data. However, in a clinical setting it may be unfeasible to position the ankle reliably in the standard pose because patients have different profiles and sizes of the thigh, shank and the calcaneus that may affect the pose of the foot/ankle complex.For measurements made on the M/L radiographs , most of the parameters were found not to be affected by rotational errors about the three axes. The only sensitive parameters were MDA and MDV, which have important implications for TAA because these parameters are related to determining the optimal cutting level of the bone and the relevant instrumentation in ankle arthroplasty in order to preserve the healthy bone of the tibia and fibula as much as possible . For meaSince it is unrealistic to expect the ankle to be positioned in the standard pose reliably in a clinical setting, one possibility is to try to reduce the 2D measurement errors given that 2D and 3D measurements of most of the morphological parameters were found to be correlated linearly. The linear regression equations obtained in the current study were shown to be effective in correcting the errors in the 2D morphometric measurements, especially for perturbation less than 3 degrees about the three individual axes. However, when the positioning errors became too large, such as up to 6 degrees, parameters sensitive to the axial perturbations, namely MDA, MDV, TaW and MLATa, were difficult to correct. This is mainly because the linear regression equations derived from the data in the standard pose were less effective in describing the 2D and 3D relationship accurately. Therefore, care should be exercised in positioning the ankle in order to minimize the positioning errors in clinical practice. With physically minimized positioning errors, the current linear regression equations would be helpful for improved estimation of the morphometric parameters for clinical purposes.In the current study, CT-based computer simulation with DRRs was preferred over the direct use of simple X-ray radiographs because the latter would need more manual involvement, increasing uncertainties and errors in the measurements during imaging. For example, an accurate positioning device that allows the angular perturbations about the three axes of the coordinate system for each specimen with different sizes and morphology would be needed. Manual identification of the bony landmarks separately on the specimen and on the X-ray image would also be required. Nonetheless, since measurements based on planar radiographs are still widely used for the diagnosis of diseases and pre-surgical evaluations of the ankle joint in orthopedic practices, it is essential to establish some knowledge of the errors or uncertainties in the radiograph-based measurements. The current results identified the parameters whose 2D measurements were sensitive to rotational pose errors during imaging. Linear regression equations were also obtained for correcting 2D measurements for clinical use. Further studies are needed to quantify the sensitivity, reliability and correction equations for the parameters when subject to errors in the ankle joint positions , as well as translational pose errors such as when the source of the X-ray is not focused on the reference points used for positioning during imaging. Further studies may also include reliability analysis under real life conditions to account for intra- and inter-observer variability that have been suggested to be the main source of artifacts in clinical practice .In the standard pose for imaging, most ankle morphometric parameters measured using 2D images were highly correlated to the gold standard values defined by the 3D CT data, with some morphological parameters not significantly different from the gold standard values. For measurements made on the M/L images, the only parameters sensitive to rotational pose errors were MDA and MDV, which had important implications for determining the optimal cutting level of the bone during TAA surgery. For measurements made on the A/P images, TaW underestimated the standard values by up to 31.2% with only a moderate reliability, suggesting that pre-surgical evaluations based on TaW should be made with caution so as to avoid inappropriate selection of the sizes of the TAA prosthesis. The linear regression equations of the relationship between 2D and 3D measurements will be helpful for correcting the errors in 2D morphometric measurements for clinical applications.The authors declare that they have no competing interests.CCK prepared the specimens, participated in the CT data collection and analysis, and drafted the manuscript. HLL developed the software programs, performed data analysis and helped draft the manuscript. TWL participated in the study design, methodological development, interpretation of the results and drafting the manuscript. CCL helped develop the software programs and interpret the results. AL participated in data interpretation and the revision of the manuscript. MYK participated in the study design and CT data collection. HCH participated in the study design and revision of the manuscript. All authors read and approved the final manuscript.


Health professionals are often at risk of accidents during the caution, and that these exposures may occur to the transmission of pathogens such as human immunodeficiency virus (HIV), Hepatitis B and Hepatitis C. The Committee on Infection Control has a fundamental role in the performance of the first visit to the accident, through guidance as to the serological monitoring and adherence to the monitoring of employees.In this study in a general hospital, were analyzed all accidents involving biological material in the period 2003 to 2010, by attending at the time of the accident, with research into the causes and serological employees exposed during the accident involving biological material and collection of serologic of source patients.During the period, 398 accidents were monitored, and 234 (59%) with negative source patients, eight (2%) with the source patient Hepatitis B, 42 (10%) source patients with hepatitis C, 20 (5 %) source patients with HIV and 90 (23%) patients with unknown source, which enables these patients have positive tests questioned. During the first three visits were diagnosed (0.75%) employees were seropositive for hepatitis C and no history of accidents with biological material. Of these, 16 (4%) did not complete the follow-up due to the shutdown of the institution, despite subsequent contact. E 382 (96%) employees did not show seroconversion.Adherence to institutional guidelines for sharps injuries and the traceability of the source, the introduction of chemoprophylaxis, immunization against hepatitis B, are shown to be key measures in the management of accidents with biological material.None declared.


The aim of this research was to compare the efficiency of different murine lines for genotoxicity assays. Rats and mice of different murine lines were used. The spontaneous and induced indexes were evaluated according to alkaline comet assay of peripheral blood leukocytes, micronucleus and chromosomic aberration assay of bone marrow cell, and sperm head morphology assay. In most of the evaluated assays the line of Balb/c mice turned out to be the ideal biomodel, with less spontaneous indexes and high induced indexes to the mutagen used; allowing to detect in a narrow error margin those substances that are classified of very low genotoxicity. These results demonstrate that genetically the line of Balb/c mice in both sexes is more stable than the other ones evaluated. This suggests the use of the Balb/c line on in vivo genotoxicity assay will increase sensibility and robustness. The CF is an alquilant agent that forms monoadducts and crossed connections between chains as consequence of the appearance of ruptures for reparative mechanisms effects. This drug is used with great effectiveness as an antineoplastic , Lewis and Wistar of both sexes in the alkaline comet assay of peripheral blood leukocytes, micronucleus and chromosomic aberrations in cells of the bone marrow and the sperm head morphology assay.To compare the mouse line and rat selected of both sexes in the alkaline comet assay of peripheral blood leukocytes, micronucleus and chromosomic aberrations in cells of the bone marrow and the sperm head morphology assay.The assessment of this research were:Lines of mice were Balb/c, NMRI, OF-1 and C57/BL6/Cenp of both sexes, 8–9 weeks of age, body weight 26–30 g at the end of the quarantine. Rats of the genetic lines SD, Lewis and Wistar were used, 6–8 weeks, 180–210 g at the end of the quarantine. Animals were randomly distributed in groups of 5 animals each, with 10 animals in total (in the two replics).®, Lemery, CORP), which was diluted in saline solution (NaCl) to 0.9%. The solution was administered immediately after being prepared, 24 than 48 hours and then at the 24 hours before the euthanasia at 10 ml/kg , group 2: Tween 65 at 2%, group 3: NaCl to 0.9%, group 4 (positive control): CF in dose of 50 mg/kg, via intraperitoneal (i.p),  Shayne,  and 52 det al., At the end of experiment we were able to do the alkaline electrophoresis of individual cells assay (comet assay), micronucleus assay in bone marrow, chromosomic aberrations assay in bone marrow and the sperm head morphology assay as previously reported  was done and counts were done with a NewBauer chamber and microscope Olympus BH-2, 10× , total of cells with structural aberrations in the chromosomes, mitotic index (number of cells that were in the phase of cellular division of metaphase) and the per cent of peripheral blood leukocytes, for levels of damage in the DNA according to level 1, 2, 3, 4 of less to more damage  and the categorical variables by Chi Squared (p<0.01) /normochromatic erythrocytes (NE), the genotoxicity index (% of PE that contain micronucleus), as well as the number of PE with 1, 2 or more than 2 micronucleus, as index of damage severity. In this line we obtained the lower spontaneous results and the highest induced results, being observed a high sensibility of this animal biomodel to detect clastogenic compound , achroleine, carboxyl-phosphamide and 4-cecophosphamide that are barely active. The CF is inactivated for microsomal and hepatic enzymes with active participation of the P-450 cytochrome (Arencibia et al., et al., For the first time it was performed a deep characterization, endorsed statistically by several biomodels keeping in mind their efficiency (Arencibia


Although microwave irradiation has been used for disinfection of dental stone casts, there are concerns regarding mechanical damage to casts during the process. The aim of this study was to evaluate the effect of microwave irradiation on the compressive strength (CS) and diametral tensile strength (DTS) of stone casts. In this in vitro study, 80 cylindrical type III and IV stone models (20 × 40 mm) were prepared and divided into 8 groups of 10. The DTS and CS of the specimens were measured by a mechanical testing machine at a crosshead speed of 0.5 cm/min after 7 times of frequent wetting, irradiating at an energy level of 600 W for 3 minutes and cooling. Data were analyzed by Student’s t-test. Microwave irradiation significantly increased DTS of type III and IV to 5.23 ± 0.64 and 8.17 ± 0.94, respectively (P < 0.01). According to the results, microwave disinfection increases DTS of type III and IV stone casts without any effects on their CS. Despite being effective procedures for disinfection, the materials and the processes involved have been shown to exert detrimental effects on the physical and mechanical characteristics of dental casts.6 In addition, further precautions restricting the use of these materials include their toxicity to humans and the environment, generation of resistant species, and complexity and sensitivity of the techniques.10Gypsum casts may cause cross-infection between the clinic and the laboratory during prosthodontic treatments.15 However, being a new method, there are not sufficient data on the effect of microwave irradiation on the physical, including surface roughness and dimensional accuracy, and mechanical characteristics of stone casts. The mechanical properties including compressive strength (CS) and diametral tensile strength (DTS) in this regard have, in particular, direct influences on fracture and abrasion resistance of dental stone casts. However, previous studies on the subject have yielded controversial results.18 Moreover, these studies have not investigated the effect of repeated irradiations, as considered in chemical disinfection studies.6To overcome these restrictions, other methods such as microwave disinfection of acrylic resins and stone casts have been proposed and assumed effective.22 One study has shown that the sterilization of stone casts can be achieved at 600 W in 3 minutes,23 an energy level which is in the safe range for stone casts. Contrary to other studies which have evaluated the effects of irradiation on mechanical properties of stone casts in one irradiation cycle, if according to the conditions of this study, it could be shown in multiple disinfection procedures by irradiation, similar to the condition of casts in the clinic and laboratory, that the DTS and CS of stone casts do not decrease, this technique can be applied as a simple and reliable disinfection technique, without the disadvantages of chemical techniques, in every stage of clinical procedures.In spite of the controversy, many studies emphasize that only low-to-medium levels of energy are safe and preserve the CS, DTS and dimensional accuracy of stone casts.The hypothesis tested in this study was that DTS and CS of stone specimens are not affected by microwave irradiation at 600 W in seven separate exposures for three minutes in each run in comparison to the control group.22 A weighed amount of powder was added to water in a rubber bowl and mixed manually to a smooth consistency and then placed in a vacuum mixer  for 25 seconds under the vacuum condition of 0.8 to 1 mm Hg to eliminate porosity; then the mixture was gently poured into the silicon mold on a vibrator. Subsequently, a glass plate was placed on the silicon mold to create a smooth and parallel surface. Forty-five minutes after mixing, the cylindrical specimens were removed from the silicon mold and weighed on a digital weighing machine. Based on similar studies,6 ten specimens were prepared for each study group; on the whole, 80 specimens were prepared with the same dimension and weight without any obvious defects at ×3 magnification in this manner and stored at ambient temperature of 23±2°C and a relative humidity of 45–50%  and type IV  dental stones were used for the purpose of this in vitro study. The water/powder ratio used was according to manufacturer’s instructions. A silicon rubber mold  with four cylindrical spaces, 20 millimeter in diameter and 40 millimeter in height, were prepared from stainless steel models according to ANSI/ADA No. 25.f 45–50% .Twenty-four hours after preparing the stone specimens, half of them were randomly separated for irradiation. In the first stage, the casts were wetted by spraying water until they did not absorb any more, but without any visible water on surfaces when they were placed in the microwave oven to prevent micro-explosion and cracking following the rapid formation and release of vapor.The specimens were irradiated in a microwave oven  for 3 minutes at 600 W. To prevent heat shock, the casts were allowed to cool in air. The sequences of wetting, irradiation and cooling were repeated seven times, while a container with 400 mL of water was placed in the chamber to protect the magnetron of the microwave oven from excess heat after all the moisture of the specimens evaporated.One week after the production of casts, a mechanical testing machine  was used for testing the irradiated and un-irradiated specimens with a load of 10 Kg at a crosshead speed of 0.5 cm/min .To evaluate the CS of the specimens, each sample was placed in a perpendicular position in the testing machine and the maximum force (MPa) for breaking was recorded for CS.To measure the DTS, each specimen was placed diagonally in the testing machine and after measuring the maximum force for breaking the specimen , the folwhere “P” is the force applied, “D” is the thickness of the specimen and “T” is the height of the specimen. The results were also recorded in MPa. The specimens that broke into more than two pieces were excluded from the study. In this study CS and DTS were measured by a specialist in material engineering, who was blind to the process of preparing the specimens. Data was analyzed by Student’s t-test using SPSS 15 software.The comparison of the CS in type III and type IV stone casts for irradiated and un-irradiated specimens are presented in The comparison of DTS in type III and type IV stone casts for irradiated and un-irradiated casts are presented in The aim of this study was to evaluate the effect of microwave disinfection on DTS and CS of type III and IV dental stones. The results showed that the DTS of both types in the group irradiated with a 600-W beam for 3 minutes was more than that in the groups that did not undergo irradiation; however, there were no differences in the CS.22 An increase in DTS in type III and IV stone casts after irradiation with an energy level of 900 W for 5 minutes was also observed later.17The results of this study are consistent with those of previous studies which have shown that the DTS in stone casts dried by microwaves with an energy level of 600 W for 10 minutes was more than that in air-dried casts 2 hours after irradiation.18 Although casts dried with microwave with maximum power for five minutes had higher surface hardness compared to air-dried casts after 4 hours, an ultimate decrease in surface hardness was observed after 24 hours.18 According to another study, an energy level of 550 W for about five minutes increases CS in type III stone casts after 24 hours.20 However, another study showed increases in CS in type III and IV stone casts at an energy level of 900 W for about 5 minutes as mentioned above.17Lower energy levels have been suggested for the process of irradiating and heating of gypsum products, especially in denser casts, in order to prevent crack formation as a result of rapid formation and release of vapor.The controversial results might be attributed to different techniques of irradiation applied; when the irradiation period at 900 W is divided into two 2.5-minute periods, the interruption in the period can decrease the temperature and prevent damage from high levels of energy.4.2H2O under energetic microwaves, resulting in dehydrated poor casts.25One of the main reasons for lower strength of oven-dried casts compared to air-dried ones is lack of water molecules in the early hours, which are necessary to form, grow, sediment and join together the crystals of CaSO25 The comparison of the strength of the specimens at this stage is useful in detecting even minor effects of microwave irradiation on casts. This fact was one of the considerations in the current study. Another consideration in the present study was the seven-time irradiation as an average number of times that casts need to be disinfected during prosthodontic procedures.6 Moreover, wetting the casts before irradiation was an attempt to imitate the condition of casts in microbiologic studies.23Additionally, in order to achieve maximum strength, the excess water of stone casts should be removed, a process which takes seven days after pouring in the ambient temperature.Studying the effects of microwave irradiation on the physical properties, such as surface roughness, detail reproduction and dimensional stability; mechanical properties such as surface hardness and abrasion resistance, and the research for techniques that increase microbicidal efficiency of microwave can be new fields of investigation.Under the limitations of this study, microwave irradiation of type III and IV stone casts at 600-W energy level for 3 minutes could enhance their DTS without affecting their CS.


According to the National Heart, Lung, and Blood Institute of the National Institute of Health, hypotension refers to an abnormally low blood pressure (BP). In childhood, hypotension can be determined according to two different definitions: BP below the 5th percentile or below two standard deviations (SDs) of the mean for age and gender . A graphWe used a computerized calculation method to develop high-resolution graphics containing curves with 5,841 points each, to depict the main percentiles associated with low BP for girls from 1 to 17 years old in the 50th percentile of height. Each point represents the calculation of the polynomial equation that includes the statistical processing of the last Report on Blood Pressure in 2004 . We alsoFive monotonic curves of diastolic BP for girls representing the 50th, 25th, 10th, 5th, and 2.275th (-2SD) percentiles were built (Figure The correction of female diastolic BP for height is of minimal significance, and this updated graphic can be used to diagnose low diastolic BP for girls. Clinical studies are necessary to determine the diastolic BP percentile that better represents clinically significant hypotension.


Defects in the DNA damage response often lead to an increased susceptibility to cancer, and so the DDR presents an interesting set of novel therapeutic targets. The maintenance of genomic integrity by the DDR has also been found to be involved in the process of organismal ageing. While the removal of cells containing damaged DNA can be beneficial in the prevention of cancer, it may contribute to both normal and pathological ageing. The repair pathway activated in res-ponse to such agents is dependent on the type of lesion generated. Base excision repair (BER) and nucleotide excision repair (NER) pathways are typically activated in response to damage to individual DNA bases , while bThe DDR comprises multiple proteins, and a complicated network of signaling pathways to ensure that the processes of DNA repair, cell cycle arrest or the triggering of the apoptotic cascade are correctly regulated. Since the individual elements of the DDR have been reviewed at length elsewhere, this paper will aim to give a brief overview of the key components in order to further discuss how the DDR can be targeted to treat cancer.ATM (ataxia telangiectasia mutated) is a member of the phosphatidylinositol 3-kinase related family of serine/threonine protein kinases (PIKKs) . ATM is Multiple proteins have been shown to be phosphorylated downstream of ATM, including the known tumor suppressor protein p53, structural maintenance of chromosomes (SMC) 1, which is known to engage the S phase checkpoint, the breast and ovarian cancer susceptibility protein BRCA1 and the checkpoint kinase Chk2 -19. ThesATR (ATM-Rad3-related) is also a member of the PIKK family, and while being related to ATM, plays a distinct role in the DDR. Loss of ATR has been shown in mouse models to convey embryonic lethality , suggestThe initial step in ATR signaling is the binding of replication protein A (RPA) to single stranded DNA (ssDNA), which recruits the ATR-ATRIP complex to the DNA damage. The recognition of neighboring DNA ends by the RAD9-RAD1-HUS1 (9-1-1) complex brings the protein TOPBP1 into the vicinity of ATR-ATRIP to stimulate ATR activation -24. As w1/S and G2/M checkpoints, and the repair of DSBs by HR .1/S arrest by BRCA1 is known to require several other proteins. The inhibition of gene transcription is regulated by the interaction of hyposphosphorylated pRB with the E2F transcription factor. This hyposphosphorylated state of pRB is thought to be maintained by its interaction with BRCA1 [The induction of Gth BRCA1 . Cell cyth BRCA1 .The intra-S phase checkpoint is also regulated by BRCA1, through the BRCA1B complex containing BRCA1, BACH1 and TOPBP1 (reviewed in ). This c2/M checkpoint. The interaction of CtIP and MRN has been found to regulate the resection of DNA ends [2/M checkpoint.The BRCA1C complex, comprising BRCA1, the carboxy-terminal binding protein interacting protein (CtIP) and the MRN complex, has an essential function in regulating the GDNA ends , a proceDNA ends ,37, ther2/M checkpoint, stimulation of BRCA1 and Chk1 expression and the interaction and recruitment of the BRCA2/ RAD51 complex for the execution of HR. The documented failure of various DDR proteins including NBS1, phosphorylated ATM and ATR to localize to sites of DNA damage in the absence of BRIT1 suggests that BRIT1 functions upstream of these proteins as a scaffold to bridge phosphorylated H2AX and the DDR .In 2009, BRIT1 was shown to recruit the SWI-SNF chromatin remodeling complex to sites of DNA damage in an ATM/ATR-dependent manner. This recruitment is thought to facilitate the unwinding of chromatin following DNA damage, thereby allowing access of the required repair proteins. This shows that BRIT1, therefore, plays a critical role in both HR and NHEJ mediated repair .The interplay between BRIT1, BRCA1 and the other upstream DDR components discussed here is illustrated in figure The checkpoint kinases Chk1 and Chk2 are widely considered to be the major effectors of the DDR in regulating cell cycle checkpoints and coordinating this with DNA repair.1/S, intra-S phase or G2/M checkpoint [As with ATR, Chk1 has found to be critical for embryonic development, suggesting that it is active at multiple stages within the cell cycle . In respeckpoint  Fig. 2)1/S, intrIn contrast, Chk2 is not considered to be vital for embryonic development, but is believed to be activated following DNA damage. Although Chk2 has been found to phosphorylate CDC25A, its role in the DDR is thought to be related more to its effect on p53 stabilization than cell cycle arrest .The frequency of p53 mutation and loss in human cancers suggests a critical role for p53 in the maintenance of genomic integrity. p53 can be phosphorylated by a number of DDR proteins, including ATM, ATR and Chk2 ,44,45. T1/S and G2/M transitions through its inhibition of cyclin-dependent kinases (CDK) [One of the key activities of p53 is thought to be the induction of expression of p21, a known regulator of both the Ges (CDK) . A furthOver 30 years ago it was established that cells within a tumor are derived from a single genetically unstable cell, and that the population as a whole continues to acquire further chromosomal abnormalities over time . HoweverMany of the DDR components including BRCA1  and BRITWhile the loss of BRCA1 has been shown to lead to the development of mammary tumors in mouse models, the genetic diversity within those tumors suggests that the loss of BRCA1 is not directly responsible for tumorigenesis. It is more likely, therefore, that the role of BRCA1 in the initiation of cancer is a result of its effects on DNA repair and the maintenance of genomic integrity . These mSimilar to BRCA1 −/− tumors, BRIT1 −/− tumors also display numerous chromosomal aberrations. Interest-ingly, BRIT1 is expressed on the short arm of chromosome 8, a region which has been found to be altered in various forms of cancer -56. A siIn addition to its role in the maintenance of genomic integrity, the DDR has been hypothesized to play a critical role in organismal ageing. Ageing, resulting from the accumulation of damage to molecules, cells, organs and tissues over time, is believed to be caused by two cellular processes: senescence and apoptosis.Senescence was first described by Hayflick and Moorhead in 1961 when it was noted that fibroblasts entered a state of permanent growth arrest following serial cultivation; a fate that was not shared by cancer cells . FurtherReplicative senescence results from the progressive shortening of telomeres with repeated rounds of cell replication. In 2007, Feldser and Greider demonstrated that the shortening of telomeres in mice was related to a suppression of tumor incidence as a result of induction of a p53-dependent senescence pathway. This p53 response to critically shortened telomeres has been demonstrated to be a result of activation of the DNA damage response (reviewed in ), since et al.[However, the role of p53 in the induction of a senescent phenotype has been debated by recent studies, which have shown that p53 can either activate or suppress senescence. The determination of cell fate has been postulated to depend on the specific transcriptional activities of p53. Several recent reports have proposed a mechanism by which p21-induced cell cycle arrest could be converted into either a quiescent or senescent phenotype, depending on the activity of p53 and its inhibition of the protein kinase mammalian target of rapamycin (mTOR) -62. Brieet al.. This pup53 is not the only DNA damage response protein to be associated with replicative senescence. Studies in mice have shown that the deletion of ATM causes an increase in both chromosomal end-to-end fusion events and cell cycle-dependent telomere loss. These events were accompanied by a premature ageing phenotype, with symptoms including increased hair graying, alopecia and marked weight loss . A furthSenescence can also be induced by the overexpression of oncogenes. This has recently been reviewed elsewhere  and so win vitro[in vivo[The accumulation of DNA damage does not necessarily lead to cellular senescence. The activation of p53 by DNA damage has been well documented, and its role in the regulation of expression of pro-apoptotic proteins has been recently reviewed . The facin vitro and in vo[in vivo in respoThe DDR is a complex network of proteins, comprising DNA damage recognition, signal transduction, transcriptional regulation, cell cycle control and DNA repair. The maintenance of the DDR is essential for faithful replication of the genome, and so is critical for cellular survival. The loss of certain DDR components can lead to an increased susceptibility to cancer due to the ensuing genomic instability and the subsequent mutation to genes required for cellular replication and division. The DDR is also involved in the induction of senescence and apoptosis when the damage cannot be repaired. While this can prolong longevity during early stages of life due to the suppression of tumorigenesis, it may become detrimental in ageing due to the loss of stem and progenitor cells for renewal. This is a phenomenon referred to as antagonistic pleiotropy, and it highlights the importance of carefully balanced cell signaling cascades and regulatory systems in the maintenance of survival. Further studies of the roles of DDR-associated proteins, along with the discovery of new ones, will therefore not only enhance our understanding of cancer and mechanisms to treat it, but will also enhance our understanding of the ageing process. This may uncover ways to treat premature ageing or other age-related pathologies, such as the decline of the immune system in the elderly.


S. cerevisiae for the first time. Our model incorporates all recently known and several proposed interactions between the osmotic stress response pathway and the cell cycle. This model unveils the mechanisms that emerge as a consequence of the interaction between the cell cycle and stress response networks. Furthermore, it characterises the role of individual components. Moreover, it predicts different phenotypical responses for cells depending on the phase of cells at the onset of the stress. The key predictions of the model are: (i) exposure of cells to osmotic stress during the late S and the early G2/M phase can induce DNA re-replication before cell division occurs, (ii) cells stressed at the late G2/M phase display accelerated exit from mitosis and arrest in the next cell cycle, (iii) osmotic stress delays the G1-to-S and G2-to-M transitions in a dose dependent manner, whereas it accelerates the M-to-G1 transition independently of the stress dose and (iv) the Hog MAPK network compensates the role of the MEN network during cell division of MEN mutant cells. These model predictions are supported by independent experiments in S. cerevisiae and, moreover, have recently been observed in other eukaryotes.The cell cycle is a sequence of biochemical events that are controlled by complex but robust molecular machinery. This enables cells to achieve accurate self-reproduction under a broad range of different conditions. Environmental changes are transmitted by molecular signalling networks, which coordinate their action with the cell cycle. The cell cycle process and its responses to environmental stresses arise from intertwined nonlinear interactions among large numbers of simpler components. Yet, understanding of how these pieces fit together into a coherent whole requires a systems biology approach. Here, we present a novel mathematical model that describes the influence of osmotic stress on the entire cell cycle of  The cell cycle is the most fundamental biological clock underlying all forms of life. It enables faithful duplication of the entire set of genes before cell division, ensuring stable cell proliferation. The cell cycle can be considered as a sequence of biochemical events governed by a complex but robust molecular network. This network has evolved in a sophisticated way, allowing cells to achieve accurate self reproduction in various conditions. Environmental changes are transmitted by molecular signalling networks that allow cells to react accordingly. Signal transduction networks, however, do not work in isolation, but coordinate their action with the cell cycle machinery; allowing flexible timing of crucial cell cycle events, adapted to the type and level of stress. In the past, cell cycle and stress response networks have generally been studied in separation. It has recently become clear, however, that to understand cellular responses to stresses, cell cycle and signalling networks have to be considered simultaneously. Recent studies, particularly in the case of osmotic stress S. cerevisiae. First modelling approaches have addressed the interaction between the osmotic stress response and the G1 phase of the cell cycle of S. cerevisiaeS. cerevisiae. The model unveils that the influence of the osmotic stress on different stages of the cell cycle attributes to interaction among many components of the entire cell cycle network rather than a single element. It also provides a tool for further investigation of the molecular processes and cell behaviour of the budding yeast cells under various environmental conditions and experimental setups.The molecular machinery, which regulates DNA replication and segregation, is highly conserved from unicellular eukaryotes to multicellular eukaryotes S. cerevisiae) is regulated by the availability of its cyclin partners, inhibitory tyrosine phosphorylation (like phosphorylation of Cdc28 by Swe1) and binding to stoichiometric CDK inhibitors (like Sic1) The cell cycle of eukaryotes consists of two main phases, the S (DNA Synthesis) phase and the M (Mitosis) phase, which are separated by the G1 (Gap1) and G2 (Gap2) phases. In the G1 phase, the cell substantially increases in size and prepares for the S phase, during which DNA replication occurs. The G2 phase provides the cell with additional time to grow and to activate regulatory mechanisms in preparation for cell division. During the M phase chromosome segregation and nuclear division take place, and the cell divides into two isogenic cells. The cell cycle has three main transitions  MAPK signalling plays a key role In untreated cells the G1-to-S transition is triggered by the activation of the Cdc28-Cln2 complex which enables Cdc28 to target Sic1 (CKI) for degradation  Before the transition to the M phase, ower see . This re(ii) Immediately after the beginning of the FINISH process, instead of having a delay, the progression of the cell cycle is accelerated  the Sic1 stabilisation mediated by Hog1PP  et al.A significant observation by Reiser cdc15 cells to osmotic stress, we reproduce the known experimental results. Our simulations show that cdc15 cells, as well as further temperature-sensitive MEN mutants, end mitosis and enter a new cell cycle after exposure to various doses of osmotic stress, as measured in the experiments  upon osmotic stress in late S or early G2/M phase, cells undergo a second incidence of DNA replication before mitosis, (ii) cells stressed at late G2/M phase have an accelerated exit from mitosis and get arrested in the next cell cycle.In non-stressed cells, DNA re-replication is prevented due to the inhibitory role of Cdc28-Clb5 in the assembly of the pre-replicative complex. When cells are osmotically stressed at the end of the S phase or early G2/M phase, however, the activity of Cdc28-Clb5 is downregulated by Hog1PP. This cancels the inhibition of Cdc28-Clb5 on the pre-replicative complex formation, and thereby causes increased activity of Cdc6 and Cdc14. Then, when Cdc28-Clb5 recovers, after Hog1PP returns to its basal level, a second incidence of DNA re-replication is initiated before mitosis. Note that the level of Cdc6 is lower than in the first incidence of DNA replication, but there is experimental evidence that low Cdc6 levels are sufficient to licence the origin of replication and transfer the cell to the S phase S. cerevisiae Cdt1 is also present, but to our knowledge, it has not been reported that it is phosphorylated by HogPP.The alternative prediction derived from our model is that Hog1PP may exert another, as yet undetected, level of control on licensing factors to prevent re-replication of DNA. Interestingly, in human cells the licensing protein Cdt1, which assists the assembly of the pre-replicative complex, is phosphorylated by the stress-activated mitogen-activated protein (MAP) kinases. Phosphorylated Cdt1 is then rapidly degraded, thereby inhibiting DNA re-replication upon osmotic stress S. cerevisiae. However, recent studies on the influence of osmotic stress on dividing leaf cells in the model plant Arabidopsis thaliana also shows early exit from mitosis, as our model predicts for budding yeast S. cerevisiae, it would strongly indicate that a key component linking the M phase and osmotic stress response is missing; our model suggests that this component should hinder the Clb2 inactivation mechanisms to inhibit accelerated exit from mitosis.The second main prediction of the model, namely the accelerated exit from mitosis, has, to our knowledge, not yet been tested experimentally for et al. for the cells which are at the G1 phase at the onset of stress Furthermore, we find that the cell cycle progression is delayed approximately linearly with stress dose if the cells are stressed at the G1, S or early G2/M phase. This linear increase of the delay with the stress dose has been experimentally observed by Adrover cdc15 cells can progress through mitosis due to increased Cdc14 activity mediated by Hog1PP et al.cdc15 cells under osmotic stress to the next cell cycle.Finally, our model provides a mechanism that explains why MEN (Mitotic Exit Network) temperature sensitive mutant cells undergo mitosis under osmotic stress. It has been suggested that Therefore, stabilisation of Sic1 by Hog1PP across all cell cycle phases, seems to be the most important biochemical event in the interaction between osmotic stress and cell cycle progression.S. cerevisiae. However, the molecular basis of control of two crucial events of the cell cycle, DNA replication and segregation, is highly conserved in higher eukaryotes, including humans, with CDK playing a universal role Arabidopsis thalianaOur mathematical model is built based on the molecular mechanisms of the model organism Note that this model has been developed for doses of osmotic stress between 0.4 M and 1 M NaCl. For higher doses of stress, other links and components, not identified yet, may be involved in cellular response to osmotic stress. Moreover, the repair mechanisms for DNA replication errors are not included in the model. Therefore this model cannot predict the recovery process for the cells that have two sets of DNA because of osmotic stress. Also note that our model is built to unveil the structure of the regulatory mechanisms of the cell in response to osmotic stress rather than to make an exact quantitative prediction of the levels of the proteins involved. In order to achieve that, further experiments and data fitting are necessary.In summary, our model provides a series of novel predictions for the interactions between the cell cycle and the osmotic stress response, which on one hand are validated by existing experimental data, and on the other hand, suggest new experiments.The construction of our model started from two basic modules: the cell cycle module and the osmotic stress response module In general the time profile of the concentration We used mass-action kinetics, Michaelis-Menten kinetics, and Hill functions to describe the production/activation and degradation/inhibition interactions of each component The set of initial conditions and parameters used in our simulation are presented in Tables S2 and S3 in et al.et al., Clotet et al. and Yaakov et al.sic1Δ cells is comparable with the known delay The parameters used in the modelling of the morphogenesis checkpoint  experiments in the case of a synchronised cell population. The cell cycle, however, is a continuous rather than a discrete progression of biochemical events. When referring to the model predictions though, it is useful to have a precise definition of the borders between the phases. Therefore, here we introduce a mathematical definition of the limits between the cell cycle phases, which we use throughout the paper: the G1 phase starts right after cell division, and finishes when the level of Cdc28-Clb5 crosses the level of Sic1 Matlab Programme S1The Matlab code is written in such a way that the dose of the stress and the time point of application of the stress can be adjusted by the user.(ZIP)Click here for additional data file.Supporting Information S1(PDF)Click here for additional data file.


This study was performed between 2007-2012 and encompasses 94 patients. The patients were divided in two groups. The first group included the patients who have benefited from surgical treatment (42 cases) and the second one included patients who benefited from conservative treatment. Out of the total number of cases, 63 cases showed an intact simple bone cyst that was most of the time an accidental discovery. 31 patients presented with fracture sustained on a simple bone cyst. There were 63 boys and 31 girls. Their mean age was 9.9 +/- 2.34 years. Single injection was performed for 49 patients; the rest had double or triple injections. The mean follow-up was 34.5 +/- 6.6 months. The procedure succeeded in obtaining healing in 77 cysts (82%). Cyst index of more than five and cortical index of less than 1 mm were significantly prone to pathological fractures and had significant poor results after treatment. Our results suggested that an autogenous bone marrow injection is a safe and effective treatment method for simple bone cysts, when compared with the surgical management, but sometimes-repeated injections are necessary. Cyst index and cortical width are good indicators for treatment outcome. Although the pathogenesis is still unknown [11], the lesion appears to be reactive or developmental rather than to represent a true neoplasm [1]. SBC consists of a solitary cavity lined by a membrane of variable thickness and filled with a clear yellow fluid [2]. It represents approximately 3% of all primary bone lesions [4].The simple bone cyst is the most common benign lytic bone lesion in childhood, mainly affecting the proximal femur and proximal humerus. The simple bone cyst (SBC) also called unicameral bone cyst is a tumor-like lesion of unknown cause, attributed to a local disturbance of the bone growth [15]. In this case, one examination of the fourth recurrence by polymerase chain reaction revealed a TP53 mutation [16]. Until now, only two SBC’s have been examined cytogenetically, revealing complex clonal rearrangements involving chromosomes 4,6.8,12,16 and 21 in one case and a translocation  in the second case [13]. About 65% of these cysts occur in teenagers and an additional 20% in the first decade of life. The vast majority of SBC are located in the proximal diaphysis of the humerus and femur, especially when they occur in patients younger than 17 years old [12]. The symptoms include pain, swelling, or stiffness at the nearest joint. (3) A pathologic fracture is often the first sign of the lesion.  In fact, this is the most common complication of the SBC and occurs in about 66% of the cases. In older patients the incidence of involvement of atypical sites, such as calcaneus, talus, and ilium rises significantly [13]. In these sites, the lesion is usually asymptomatic and is discovered by accident. The SBC is more common in males 3:1 and is usually detected during the first 2 decades of life [Fig. 1). Epiphyseal extension is unusual [8]. Radiographically, the appearance of an SBC is that of a centrally located, well-circumscribed, radiolucent lesion with sclerotic margins. The lesion is located within the diaphysis or the metaphysis of a long bone, abutting or being remote from the cartilaginous growth plate . The effectiveness of this method was studied according to different factors that affect the outcome including the aggressiveness of the cyst.Fig. 2). From 2007 to 2012, 94 consecutive patients with diagnosis of SBC were treated in the authors’ institute by: aspiration and percutaneous ABMI or steroid injection (methylprednisolone), and surgical treatment. Confirmation of the diagnosis was based on characteristic radiographic appearance and histological study of the specimen taken by aspiration. There were 63 boys and 31 girls. The mean age at the outset of treatment was 9.9 +/- 2.34 years (range 6-17). The cyst localizations were humerus 50 cases, radius 5 cases, V-th metacarpal 1, femur 25, peroneus 8, tibia 4, calcaneus 1 .55 cysts were uniloculated and 39 were multiloculated. Cyst size is defined as the ratio of the length of the cyst to the width of the adjacent physis, which allows a direct comparison of the sizes without requiring correction factors for different ages. The mean size of all cysts in this study was 1.857 +/- 0.473 (range 0.9-3.1). The fracture risk was evaluated by measuring the cyst index of Kaelin and Mc Ewan and the cortical thickness. The cyst index is the cyst area divided by the squared diaphyseal diameter. One or more trapezoids are drawn around the cyst area. Mechanically, a cyst is considered not prone to fracture when the index is lower than 3 and the cortical width is lower than 2 mm. The patients were divided by Cyst Index (CI) as it follows: upper limb with a CI >4 – 40 patients, upper limb with a CI <4 – 22 patients, lower limb with a CI <2 – 7 patients, lower limb with a CI 2-3,5 – 11 patients, lower limb with a CI > 3,5 – 20 patients .Surgical treatment consisted in elastic reinforcement 5 cases, dyaphisectomia + bone graft 4 cases, curettage and bone grafting 11 cases, curettage of cystic cavity and osseous substitute 8 cases, curettage of the cystic cavity and structuralized bone graft plus stabilization 24 cases . In the quantification of the results, we considered the occurrence of the recidivated cysts, time span until the total healing and the number of injections needed in the case of steroid injection and ABMI. In the case of surgical management, we encountered recidivated cysts in 14 patients. Upper limb accounted for 12 recidivated cysts and the lower limb for only 2 . Time span from the surgical intervention to total healing of the cyst varied between 6 months and 2 years with a peek incidence between 12 and 18 months (30 patients)  occurred in the cases with latent cysts, and for the cases which needed prolonged periods of time for healing several surgical procedures were needed.Fig. 7).  From the conservative point of view, in the case of steroid injection, the quantification of the results was also based on the number of injections to achieve healing. According to the number of injections until healing, patients were distributed as it follows 1 injection -0 cases, 2 injections- 2 cases, 3 injections 10 cases, 4 injections – 14 cases. Five patients had no response to the treatment whatsoever, and they were managed surgically (We used ABMI to 12 of our patients. The first and most important conclusion was that most of the patients (9) had a good response since their first injection. According to case partition and taking into account the time span from the debut of the conservative treatment and the mechanical healing of the cyst , we observed that most of the patients were grouped in the 6 months-1 year interval. The cases that needed more time for the healing were most of them part of the cast immobilization group. Fig. 8b).After the surgical management, a large proportion of the patients sustained a recidivated cyst. These recidivated cysts were encountered in those cases that were treated with curettage and osseous graft or osseous substitute .After a brief period of time that mimed healing, we encountered the reappearance of the cyst, most of the time in the inferior pole of the old cyst. In the upper limb most of the recidivated cyst appeared in the humerus. The only surgical management that did not lead to a recidivated cyst was elastic reinforcement and osseous reconstruction with diaphysectomy. The only impediment related to simple elastic reinforcement was the period of time needed for the healing of the lesion, which proved to be very long. In the case of osseous reconstruction with diaphysectomy, this procedure is prone to a far greater morbidity rate. We recommend this kind of treatments only in the cases of conservative management failure and in cases of displaced pathological fracture. The conservative treatment consisted in simple cast immobilization of the pathological fracture localized in the upper limb and without fragment displacement, ABMI or steroid injections. The last two methods of treatment were employed only in those cases that sustained a cyst without a pathological fracture or after the healing of a previous pathological fracture, which did not lead to cyst healing. Those cases that sustained a pathological fracture localized at the level of the upper limb were successfully treated by conservative means, the result being the healing of the fracture and subsequently of the cyst. In the case of the steroid injection, we encountered 5 cases that had no response, in comparison with ABMI, in which case we encountered a 100% rate of lesion healing, even from the first injection (- A recidivated cyst frequently follows surgical management by the method of curettage and grafting. - Diaphysectomy and osseous graft leads to a hundred percent healing, but it is followed by great rates of morbidity and it is unnecessarily laborious. - The logical treatment for SBC, that sustained a pathological fracture, as well as in the case of an ordinary fracture.  - Steroid injection may be accompanied by failure, and only exceptionally, healing occurs before the third injection. - ABMI is a simple cost free method, effective in both active cyst and inactive cyst, and certain signs of healing follow it in most of the cases since the first injection.


Firstly, clusters such as fibres and helices composed of C60 molecules are produced via self-assembly in supercritical carbon dioxide. However, C60 molecules are so weakly bonded to each other in the clusters that the clusters are broken by the irradiation of electron beams during scanning electron microscope observation. Secondly, UV photons are irradiated inside a chamber in which air is filled at 1 atm and the above clusters are placed, and it was found that the clusters are reinforced; that is, they are not broken by electron beams any more. C60 molecules located at the surface of the clusters are oxidised, i.e. C60On molecules, where n = 1, 2, 3 and 4, are produced according to time-of-flight mass spectroscopy. It is supposed that oxidised C60 molecules at the surface of the clusters may have an important role for the reinforcement, but the actual mechanism of the reinforcement of the clusters has not yet been clearly understood and therefore is an open question.We carry out two experiments: (1) the formation of clusters composed of C Secondly, the clusters formed on the silicon plate were moved to another chamber made of stainless steel for irradiation of UV light  for 24 h [60 molecules were so weakly bonded to each other that the clusters were broken by electron beams during the SEM observation , that is, C60 molecules to which oxygen atoms are bonded, were detected, but neither C120 nor C60-O-C60 molecules were found. In other words, the reinforced clusters were not polymerised via a chemical bond. Note that when the chamber was vacuumed and UV light was irradiated, the clusters were broken as mentioned, but dimers such as C108, C110, C112, C114, C116 and C118 were created  located at the surface of the clusters, in the reinforcement process in detail, carrying out quantum mechanical calculations. We will also be measuring the electric, electronic, mechanical and thermal properties of the fibres and helices so that the clusters may be utilised for the development of nano electron devices, nano/microelectromechanical systems and micro-total analysis systems.We will be investigating the mechanism of the reinforcement of the clusters, that is, the role of oxidised C60 molecules were placed in supercritical carbon dioxide (36.0°C), and it was found that fibres, fibre networks and helices composed of C60 molecules were self-assembled. Since C60 molecules in the clusters were bonded to each other via van der Waals interactions [60 molecules at the surface of the clusters were oxidised, and C60On molecules were created. The clusters were not broken by the electron beams any more. It is supposed that C60On molecules located at the surface of the clusters may have an important role in the reinforcement process, but the actual mechanism of the reinforcement of the clusters has not yet been clearly understood and therefore is an open question.We carried out two experiments: (1) Crystals composed of Cractions , the cluThe authors declare that they have no competing interests.SK designed the study and carried out the experiment. TF participated in the design of the study and performed SEM observation and mass spectroscopic analysis. YS participated in the reinforcement experiment. TM participated in the design of the study, coordinated the study and wrote the manuscript. All authors read and approved the final manuscript.Supplementary materials. Supplementary Material 1 - SEM observation of clusters composed of C60 molecules which were self-assembled in supercritical carbon dioxide. The accelerating voltage of electron beams is 1.0 kV. The clusters are broken during the SEM observation. Supplementary Material 2 - SEM observation of clusters. C60 molecules located at the surface of the clusters were oxidised. The accelerating voltage of electron beams is 1.0 kV. The clusters are not broken any more during the SEM observation.Click here for file


Trauma, in addition to mortality and disability experienced by an individual, imposes direct and indirect economic and social costs on a community. Traditionally, trauma is a disease of young and middle age adults, an age group which is known to be the most dynamic and economically productive of the community. Increasing our knowledge concerning the etiology and patterns of trauma seems to be the most profitable and accessible way to prevent injuries of this nature.This study was designed to evaluate the epidemiology of adult trauma in Kashan, Iran.The current study used a retrospective cross-sectional approach, enrolling all trauma adults (20 - 60 y) admitted to the Shahid Beheshti Hospital, Kashan, between 2007 and 2011. Age, gender, place of residence, work status, educational level, urban/rural location of the accident, method of transportation to hospital, injured body areas of the victims and therapeutic interventions, were extracted from the data registry and analyzed through descriptive statistics using SPSS software.A total of 22 564 patients were included in this study. Mean age of the victims was 33.18 ± 10.90 years and the male/female ratio was 4:1. Most of the victims were manual workers (61%), and they had completed primary and junior high school level education (49.4%), they were also more likely to be residents of urban areas (88.6%). Regarding the place of injury, most accidents occurred on city streets (43.8%). Approximately 40% of the total victims were transferred to the hospital by emergency medical services (EMS). During the study period, 260 deaths were recorded and among these, 76% were related to traffic accidents.Regarding the high prevalence of trauma found in manual workers with low educational levels and motorbike users, the establishment of an integrated program aimed at improving public knowledge on the use of safety and protective measures in work environments should be implemented. The use of safety protective equipment by cyclists, motorbikers and car passengers should also be enforced. Trauma is the sixth leading cause of death and the fifth leading cause of moderate and severe disability worldwide, it accounts for 10% of all mortalities . AccordiThis study was designed to describe the characteristics of injury in adult trauma patients (20-60 y) who were admitted to the Shahid Beheshti Hospital during a five year period. It is hoped that this study will provide an insight into the epidemiology of adult trauma in one of the central Iranian cities, with a population of approximately 400,000.The present study has been approved by the Research Deputy of the Kashan University of Medical Sciences and the Trauma Research Center Committee. The design was retrospective and the study population comprised all trauma patients aged 20-60 year who were admitted to the Emergency Department of the Kashan Shahid Beheshti Hospital during a period of 5 years (2007-2011). After the establishment of the Trauma Research Center in this university (2003), all information about trauma patients and casualties was entered into a computer using a database from SPSS, Version 16. To perform the present study, the information extracted from the database were; age, sex, work and educational level of the patients, injured body area, mechanism, type and Place of accident, transportation to hospital, patients’ residency , type of treatment performed on the patients (outpatient or inpatient) and the cost of treatment.A total of 22 564 trauma patients were included in the study. Demographic data are presented in Trauma is a major cause of mortality and morbidity in young and middle age adults 20 – 60 y). This age group is considered to be the most economically productive and constitutes the majority of Kashan’s households. The adult population of Kashan accounts for 77% n = 314, 394) of the region’s total population . In this present study, the average age at the time of injury was 33.18 ± 10.9 which is similar to results from other studies conducted in Iran (, 394 of 0 y. ThisAccording to this five year study, 40% of the victims were transported to the hospital by EMS. Patient transportation by ambulance increased from 11% in 2004 to 22% in 2009 in Kashan City. Coverage of ambulance transportation in Iran is 7.5% overall -14 and t


To the Editor: In 2002, the risk of transmission of epidemic typhus in the state of Mexico was assessed by analyzing serum specimens from 393 residents of previous typhogenic areas for immunoglobulin (Ig) G antibodies against Rickettsia prowazekii. Louseborne typhus has been a historic scourge in Mexico. In 1576, in a population of 9 million, 2 million deaths were attributed to epidemic typhus  in the indigenous population of the state of Mexico was 100%; in 1988, 58%; and in 1990, 15%. In 1999, indices of infestation between 5% and 12% were detected in this population ; from 1939 to 1943, 1,220 cases were reported with 707 deaths ; and from 1959 to 1963, 64 cases were reported with 14 deaths  . The prevalence of antibodies to R. prowazekii increased with age; 1–14 years of age (seropositivity 0%), 15–24 years (14%), 25–44 years (17%), 45–64 years (24%), and >65 years (48%). Thirty-three (45%) of the serum specimens had a titer of 64, 25 (34%) had a titer of 128, and 16 (22%) had a higher titer. All eight serum specimens with a titer of >512 were from persons >45 years of age.In 2002, personnel from the office of the Secretary of Health of the State of Mexico evaluated the risk to the population who lived in previously typhogenic areas to measure the impact of their programs (>65 years with a titer of 1,024 and four subjects aged >45 years with a titer of 512 suggests reactivation of latent R. prowazekii with a resulting boost in their antibody titers. These possible cases of Brill-Zinsser disease were likely not severely ill and recovered either with antimicrobial treatment that was effective against R. prowazekii or by immune control of the infection without a specific diagnosis.The high seroprevalence suggests that this population had extensive exposure to the agent of typhus and its louse vector in the past. The finding of two subjects aged R. prowazekii are absent in young persons suggests that this rickettsia has not been circulating in this population during recent years. The high seroprevalence suggests a human reservoir of latent R. prowazekii in this population. The presence of human body lice in this population indicates that there is risk of spreading R. prowazekii from an index patient with Brill-Zinsser disease to persons in contact with the patient.That IgG antibodies against R. prowazekii by scientists, physicians, and public health agencies would lead one to believe that typhus has been eliminated as a public health concern, the recent occurrence of a large epidemic in Burundi (Although the general lack of attention to


There is evidence that rapid weight gain during the first year of life is associated with overweight later in life. However, results from studies exploring other critical periods for the development of overweight are inconsistent.The objective was to investigate BMI development to assess at what ages essential differences between normal weight and overweight children occur, and to assess which age intervals the most strongly influence the risk of overweight at 8 years of age.Longitudinal weight and height data were collected by annual questionnaires in a population of 3963 children participating in the PIAMA birth cohort study. BMI and BMI standard deviation scores (SDS) were calculated for every year from birth until 8 years of age. BMI, BMI SDS and BMI SDS change in each 1-year-age interval were compared between children with and without overweight at 8 years of age, using t-tests, logistic regression analysis and the analysis of response profiles method.At 8 years of age, 10.5% of the children were overweight. Already at the age of 1 year, these children had a significantly higher mean BMI SDS than normal weight 8-year-olds, (0.53 vs 0.04). In each 1-year-age interval the change in BMI SDS was significantly associated with overweight at 8 years with odds ratios increasing from 1.14 (95% CI 1.04–1.24) per 1 SDS increase at 0–1 year to 2.40 (95% CI 2.09–2.76) at 7–8 years.At every age, starting already in the first year of life, a rapid increase in BMI SDS was significantly associated with overweight risk at the age of 8 years. There was no evidence for a specific critical period for the development of overweight. Prevention of overweight should start early in life and be continued with age-specific interventions throughout childhood. Overweight and obesity in Dutch children have markedly increased in the last decades Prevention of overweight in childhood is high on the public health agenda, but the most promising windows of opportunity for effective intervention need to be identified Several studies have addressed the question of critical time windows for overweight development. There is now consistent evidence that growth during the first year of life is associated with overweight later in childhood, adolescence or adulthood Previous analyses of the PIAMA birth cohort data suggested that BMI at 1 year of age was associated with overweight at 7 years of age In the present study, we studied BMI development of children with and without overweight at 8 years of age, and investigated which periods in childhood most strongly influenced the risk of overweight at 8 years of age. In contrast to earlier studies, we used regular 1-year-age intervals from birth until 8 years of age, which made it possible to study the influence of the first year of life as well as the subsequent 1-year-age intervals during childhood on the development of overweight at 8 years of age in the same way.The Medical Ethical Committees of the participating institutes  approved the study. Parents, carers or guardians gave written informed consent on behalf of all the minors/children involved in the study.In 1996, the large multicentre birth cohort study Prevention and Incidence of Asthma and Mite Allergy (PIAMA) was set up around three regions north , central (provinces Utrecht and Gelderland) and south-west  in the Netherlands. The aim of this cohort study was to investigate the influence of different environmental and life style factors on the development of asthma and allergic disease, and to study the possibility of asthma/allergy prevention in children by the use of mite impermeable mattress covers. At baseline this cohort consisted of 4146 pregnant women, but the study started of with 3963 newborn children because 183 (4.4%) were lost to follow-up before any data of the child had been collected. Questionnaires containing questions on asthma, rhinitis and eczema as well as various questions on lifestyle factors  were sent to the participants at 3 months of age, and annually from 1 to 8 years of age. Further details of the design of the PIAMA study have been published previously 2) was calculated from the interpolated weight and height data at each time point. Age and sex-specific cutoff points for BMI from the IOTF definition Data on weight, height and date of measurement were derived from the annual questionnaires. Parents were asked to copy their child’s weight and height data and the date of measurement from their child’s medical record, or to measure their child’s weight and height, and indicate the date of measurement themselves. To facilitate longitudinal analysis of response profiles the reported weight and height data were linearly interpolated to fixed time points. This was done for measurements between boundaries of 6 months before and after the fixed time point, which were birth, 3 months, and 1, 2, 3, 4, 5, 6, 7 and 8 years of age. The interpolated weight and height data was checked for physically impossible values, which were removed. BMI  of the baseline population was lost to follow up. Of the remaining children, 90% (3270) completed the 8-year questionnaire. Data on both weight and height at the same time points was available for 2958 (birth), 2287 (3 months), 2787 (1 year), 2761 (2 years), 2037 (3 years), 1944 (4 years), 1867 (5 years), 1938 (6 years), 2151 (7 years) and 2056 (8 years) children. BMI at all 10 time points was available for 489 out of the baseline population of 3963 children; 2616 children had at least 5 BMI measures. Loss-to-follow-up and missingness in the PIAMA study are unlikely to be completely at random. We used the multiple imputation method to avoid bias due to selective missing of data The main outcome of interest was overweight or obesity at 8 years of age. Longitudinal analyses of response profiles were conducted to compare BMI and BMI SDS development from birth to 8 years of age for normal weight and overweight children at 8 years of age. The analysis of response profiles method consists of models that contain a group (overweight at 8 yes/no) and time (age) variable, and an interaction term between group and time. The model takes into account that the repeated BMI measures within one subject are correlated Furthermore, we compared the mean BMI SDS change between subsequent time points for normal weight versus moderately overweight and obese children at 8 years of age by means of t-tests.This provides insight in the age intervals of BMI change that are most strongly associated with being overweight at age 8. Children with normal BMI development are expected to have a stable BMI SDS over time. A large increase in BMI SDS means that a child is gaining BMI more rapidly than expected and may become overweight or obese. Logistic regression analyses were conducted to investigate which periods most strongly influenced the risk of overweight at 8 years of age. Associations between BMI SDS at consecutive ages and overweight at 8 years of age were calculated, as well as associations between BMI SDS change between consecutive 1-year-age intervals and overweight at 8 years of age.Questionnaire data from the follow-up at 3 months of age and the annual follow-ups from 1 to 8 years of age were obtained for 3943, 3818, 3740, 3694, 3563, 3518, 3473, 3374 and 3270 children, respectively. Characteristics of the study population for the available case and the multiple imputed dataset (N = 3963) are shown in 2) increased rapidly during the first year, decreased until the age of 5 and slowly increased after the age of 5. Prevalence of moderate overweight and obesity was relatively stable at all ages and ranged from 7.2 to 9.1% and from 1.2 to 1.5% respectively. 2 at birth to 4.32 kg/m2 at 8 years of age, and became statistically significant from the age of 1 year onwards population of the PIAMA cohort. These measured weight and height data at 4 and 8 years of age were compared with the parental reported data at the same age. Mean differences between measured and reported weight and height data were small, but parents of children with BMI in the highest quartile underreported weight by about 0.5 kg and overreported height data by about 0.5 cm Another aspect of validity of weight and height data is that they were not measured at exactly the same age in each child. Linear interpolation of the weight and height data to fixed time points could have lead to measurement error, yet it provided the opportunity to multiple impute missing data and to apply the longitudinal analysis of response profiles method Compared to other studies the prevalence of overweight and obesity in the PIAMA cohort is relatively low. The main reason for this is that the prevalence of overweight and obesity in the Netherlands is lower than in most other Western countries. In 1997, the Dutch nationwide growth study reported a prevalence of overweight (including obesity) of 10.2% for 8-year-old boys and 14.8% for girls, while the prevalence of obesity was 1.2% and 2.3% respectively Several earlier studies have investigated the relation between increased weight gain during specific periods and the development of overweight. A certain growth period is referred to as a critical period if changes within this period increase the risk of overweight later in life Comparison of our results to the results of other studies exploring critical periods for overweight is difficult because of differences in length of follow-up, length of time intervals and age at which overweight is defined. The Barry Caerphilly Growth study in the UK showed that weight at 5 years of age, or increased weight velocity from 1 year and 9 months to 5 years were the most important predictors for BMI, waist circumference and sagittal abdominal diameter at adult age The results of the present study are in line with the results cited above, but show that the relation cannot be attributed to a specific period but rather are the result of accumulating periods starting form the first year of life. We investigated BMI SDS change during 8 subsequent year-to-year intervals from birth onwards, and did not find evidence for a specific critical period for the development of overweight. At every age, starting already in the first year of life, a rapid increase in BMI SDS was significantly associated with overweight risk at the age of 8 years. In a previous study in the PIAMA population we identified a number of early life predictors of childhood overweight, including parental BMI and birth weight


Caloric restriction, that is limiting food intake, is recognized in mammals as the best characterized and most reproducible strategy for extending lifespan, retarding physiological aging and delaying the onset of age-associated diseases. The aim of this mini review is to argue that p53 is the connection in the abilities of both the Sirt-1 pathway and the TOR pathway to impact on longevity of cells and organisms. This novel, lifespan regulating function of p53 may be evolutionarily more ancient than its relatively recent role in apoptosis and tumour suppression, and is likely to provide many new insights into lifespan modulation. Caloric restriction (CR), that is limiting food intake, is recognized in mammals as the best characterized and most reproducible strategy for extending lifespan, retarding physiological aging and delaying the onset of age-associated diseases . RestricThus despite impressive progress in identifying the key components of the CR pathway, many proximal effectors of CR induced longevity remain unknown to date. This can be related to the fact that CR causes a myriad of transcriptional and physiological changes that are related to its positive effects on lifespan -12. Are The aim of this short review is to argue that CR is indeed a regulated process and that its main regulator pathways, mTOR and Sirt-1, are connected to the p53 pathway.The CR-driven reduction of metabolic rate involves downregulation of the nutrient sensor mammalian target of rapamycin (mTOR), the evolutionarily conserved serine/threonine protein kinase that is strongly involved in most cellular functions and implicated in stimulating cell growth -21. The The absence of glucose in the cell also increases the levels of AMP, a coactivator of AMPK. Active AMPK positively regulates the activity of the TSC1–TSC2 complex by phosphorylating the TSC2 protein (resulting in the opposite activity to the AKT-1 phosphorylation of TSC2), which then turns off the RHEB G-protein and reduces TOR activity .The TOR kinase regulates two processes that can account for the observed effect of dietary restriction on longevity: translation of selected mRNAs in the cell and autophagy. The first is the rate of protein synthesis, which is modulated by the effect of TOR on the ribosomal protein S6 kinase (S6K) and on the translation initiation factor 4E-binding protein (4E-BP) . It is tBut which pathway is indispensable for prolonged lifespan by CR, inhibiting TOR or activating sirtuins? Evidence has emerged that sirtuins and mTOR are involved in the same longevity pathway . ImportaThe p53 protein and its encoding gene were first identified in 1979 because of its association with cancer -141 and The existence of p53 in short lived organisms that do not develop adult cancers, such as flies and worms, suggests that tumour suppression is not the only or, indeed, the original function of p53. Indeed, recent studies have shown that p53 and its family members, including p63 and p73, all play important roles in reproduction ,171. Emep53 interacts with IGF-1, TOR and Sirt-1 pathways, the critical pathways that regulate aging and longevity ,175 Fig. There aHere, we have reviewed the role of caloric restriction in longevity and argued that p53 is the connection in the abilities of both the Sirt-1 pathway and the TOR pathway to impact on longevity of cells and organisms. Furthermore the integration of the p53 pathway with the IGF-1 and TOR pathways brings together a number of overlapping concepts that play a central role in life processes. Through the transcriptional regulation of different target genes  p53 negatively regulates the insulin/IGF-1 and TOR signaling, creating an interpathway network that permits cells to inhibit cell growth and division to avoid the introduction of errors during these processes under stress conditions (Figure This novel, lifespan regulating function of p53 may be evolutionarily more ancient than its relatively recent role in apoptosis and tumour suppression, and is likely to provide many new insights into lifespan modulation.