Advances in surface design and biomedical applications of magnetic nanoparticles.

Despite significant efforts by scientists in the development of advanced nanotechnology materials for smart diagnosis devices and drug delivery systems, the success of clinical trials remains largely elusive. In order to address this biomedical challenge, magnetic nanoparticles (MNPs) have gained attention as a promising candidate due to their theranostic properties, which allow the simultaneous treatment and diagnosis of a disease. Moreover, MNPs have advantageous characteristics such as a larger surface area, high surface-to-volume ratio, enhanced mobility, mass transference and, more notably, easy manipulation under external magnetic fields. Besides, certain magnetic particle types based on the magnetite (Fe3O4) phase have already been FDA-approved, demonstrating biocompatible and low toxicity. Typically, surface modification and/or functional group conjugation are required to prevent oxidation and particle aggregation. A wide range of inorganic and organic molecules have been utilized to coat the surface of MNPs, including surfactants, antibodies, synthetic and natural polymers, silica, metals, and various other substances. Furthermore, various strategies have been developed for the synthesis and surface functionalization of MNPs to enhance their colloidal stability, biocompatibility, good response to an external magnetic field, etc. Both uncoated MNPs and those coated with inorganic and organic compounds exhibit versatility, making them suitable for a range of applications such as drug delivery systems (DDS), magnetic hyperthermia, fluorescent biological labels, biodetection and magnetic resonance imaging (MRI). Thus, this review provides an update of recently published MNPs works, providing a current discussion regarding their strategies of synthesis and surface modifications, biomedical applications, and perspectives.

Measuring the viscoelastic relaxation function of cells with a time-dependent interpretation of the Hertz-Sneddon indentation model.

The Hertz-Sneddon elastic indentation model is widely adopted in the biomechanical investigation of living cells and other soft materials using atomic force microscopy despite the explicit viscoelastic nature of these materials. In this work, we demonstrate that an exact analytical viscoelastic force model for power-law materials, can be interpreted as a time-dependent Hertz-Sneddon-like model. Characterizing fibroblasts (L929) and osteoblasts (OFCOLII) demonstrates the model's accuracy. Our results show that the difference between Young's modulus EY obtained by fitting force curves with the Hertz-Sneddon model and the effective Young's modulus derived from the viscoelastic force model is less than 3%, even when cells are probed at large forces where nonlinear deformation effects become significant. We also propose a measurement protocol that involves probing samples at different indentation speeds and forces, enabling the construction of the average viscoelastic relaxation function of samples by conveniently fitting the force curves with the Hertz-Sneddon model.

Subsampled digital holographic image reconstruction by a compressive sensing approach.

We show an in-line digital holographic image reconstruction from subsampled holograms with resolution improvement and lensless magnification with high noise immunity by a compressive sensing approach. Our method treats the sensed field as subsampled, low-pass filtered and projected on a Fresnel-Bluestein base in an inverse problem approach to image reconstruction with controlled lensless magnification. So, we have demonstrated by simulation and experimental results that the approach can reconstruct images with quality even when used in holograms obtained from unusual subsampling schemes.

Association of spontaneous abortion and Ureaplasma parvum detected in placental tissue.

Spontaneous abortion is considered a public health problem having several causes, including infections. Among the infectious agents, bacteria of the vaginal microbiota and Ureaplasma parvum have been associated with abortion, but their participation needs to be further elucidated. This study aims to evaluate the influence of Mollicutes on the development of spontaneous abortion. Women who underwent spontaneous abortion and those with normal birth (control) were studied. Samples of cervical mucus (CM) and placental tissue were collected to identify Mollicutes using the quantitative polymerase chain reaction methodology. Eighty-nine women who had a miscarriage and 20 women with normal pregnancies were studied. The presence of Mollicutes in placental tissue increased the chance of developing miscarriage sevenfold. The prevalence of U. parvum in women who experienced spontaneous abortion was 66.3% in placental tissue. A positive association was observed between the detection of U. parvum in samples of placental tissue and abortion. There was a significant increase in microbial load in placental tissue for M. hominis, U. urealyticum and U. parvum compared to the control group. Detection of U. parvum in CM in pregnant women can ascend to the region of the placental tissue and trigger a spontaneous abortion.

Double power-law viscoelastic relaxation of living cells encodes motility trends.

Living cells are constantly exchanging momentum with their surroundings. So far, there is no consensus regarding how cells respond to such external stimuli, although it reveals much about their internal structures, motility as well as the emergence of disorders. Here, we report that twelve cell lines, ranging from healthy fibroblasts to cancer cells, hold a ubiquitous double power-law viscoelastic relaxation compatible with the fractional Kelvin-Voigt viscoelastic model. Atomic Force Microscopy measurements in time domain were employed to determine the mechanical parameters, namely, the fast and slow relaxation exponents, the crossover timescale between power law regimes, and the cell stiffness. These cell-dependent quantities show strong correlation with their collective migration and invasiveness properties. Beyond that, the crossover timescale sets the fastest timescale for cells to perform their biological functions.

Correction: DUB3 and USP7 de-ubiquitinating enzymes control replication inhibitor Geminin: molecular characterization and associations with breast cancer.

The final sentence of the Acknowledgements should be as follows: This work was supported by grants from Instituto de Salud Carlos III (BA15/00092), Spanish Ministry of Economy and Competitiveness/EU-ERDF (SAF2016-80626-R, SAF2013-49149-R, BFU2014-51672-REDC), Fundación CajaCanarias (AP2015/008) to RF, and the Australian National Health and Medical Research (NHMRC program grant to SRL and KKK (APP1017028).

Interval colorectal cancer after colonoscopy. / Cáncer colorrectal de intervalo después de colonoscopia.

INTRODUCTION AND AIMS: Interval colorectal cancer (iCRC) can occur due to missed lesions or to a newly developed lesion. The present study aimed to assess the iCRC rate and its characteristics in our population and find possible explanations. MATERIALS AND METHODS: A retrospective study was conducted on patients with colorectal cancer (CRC) diagnosed between January 2011 and January 2015 at our department. Demographics, endoscopic data, and tumor characteristics (location, histology, staging) were collected. We identified patients diagnosed with CCR who underwent colonoscopy at our department in the previous 10years and presented the disease (iCRC) before the date of their next recommended exam. The cases of iCRC were characterized and compared with other CRC cases. Possible explanations for the appearance of iCRC were analyzed. RESULTS: A total of 266 patients presented with CRC, 61.7% were men, and mean patient age was 70.7years. We identified 10 patients with iCRC: 6 were men, and mean patient age was 71.1years. Mean time for iCRC diagnosis after index colonoscopy was 3.5±1.84years. Tumor was located in the right colon in 50% of the patients with iCRC and in 24.5% of the patients without iCRC (P=.091). More patients with iCRC had a family history of CRC (50%) than the patients with reference CRC (3.1%) (P=.000). CONCLUSIONS: In our case series, 3.76% of all CRC were iCRC. There were no statistically significant differences between patients with or without iCRC, with the exception of family history of CRC.

Ultrafast sonochemistry-based approach to coat TiO2 commercial particles for sunscreen formulation.

TiO2 is a common inorganic filter used in sunscreens due to its photoprotective effect on the skin against UV radiation. However, the use of this kind of material in cosmetics is limited by its inherent photocatalytic activity. It is known that coating on TiO2 surface can improve some features. Although, many of the methodologies used for this purpose are still laborious and time-consuming. Thus, this work reports a novel, easy, cheap and fast strategy to coat TiO2 particles by using a sonochemistry approach, aiming to decrease photocatalytic activity and to enhance colloidal stability. For this proposal, SiO2, Al2O3, ZrO2 and sodium polyacrylate (PAANa) were used to tune the surface of commercial TiO2 particles and they were applied in a sunscreen formulation. The samples were characterized by XRPD, FT-IR, DLS, EDS, SEM and TEM. The photocatalytic activity and UV-shielding ability were also evaluated. The sunscreen formulations were prepared and characterized by zeta potential, DLS, and Sun Protection Factor (SPF). FT-IR, EDS, and charge surface of the particles confirmed the success of the sonochemistry coating. Additionally, TiO2@Al2O3, TiO2@SiO2 and TiO2@PAANa show a lower photocatalytic activity than original TiO2 with similar UV-shielding ability. The sunscreens produced with the coated TiO2 have similar SPF to the one with commercial TiO2. Specifically, the sunscreen with TiO2@PAANa shows an increase in colloidal stability. Herein, the incorporation of the sonochemical-coated TiO2 particles in sunscreen formulations may produce sunscreens with better aesthetic appearance and a greater health security due to its lower free radicals production.

DUB3 and USP7 de-ubiquitinating enzymes control replication inhibitor Geminin: molecular characterization and associations with breast cancer.

This corrects the article DOI: 10.1038/onc.2017.21.

DUB3 and USP7 de-ubiquitinating enzymes control replication inhibitor Geminin: molecular characterization and associations with breast cancer.

Correct control of DNA replication is crucial to maintain genomic stability in dividing cells. Inappropriate re-licensing of replicated origins is associated with chromosomal instability (CIN), a hallmark of cancer progression that at the same time provides potential opportunities for therapeutic intervention. Geminin is a critical inhibitor of the DNA replication licensing factor Cdt1. To properly achieve its functions, Geminin levels are tightly regulated through the cell cycle by ubiquitin-dependent proteasomal degradation, but the de-ubiquitinating enzymes (DUBs) involved had not been identified. Here we report that DUB3 and USP7 control human Geminin. Overexpression of either DUB3 or USP7 increases Geminin levels through reduced ubiquitination. Conversely, depletion of DUB3 or USP7 reduces Geminin levels, and DUB3 knockdown increases re-replication events, analogous to the effect of Geminin depletion. In exploring potential clinical implications, we found that USP7 and Geminin are strongly correlated in a cohort of invasive breast cancers (P<1.01E-08). As expected, Geminin expression is highly prognostic. Interestingly, we found a non-monotonic relationship between USP7 and breast cancer-specific survival, with both very low or high levels of USP7 associated with poor outcome, independent of estrogen receptor status. Altogether, our data identify DUB3 and USP7 as factors that regulate DNA replication by controlling Geminin protein stability, and suggest that USP7 may be involved in Geminin dysregulation during breast cancer progression.

Physiological strain to prolonged exercise bouts at the walk-run transition speeds depends on locomotion mode in healthy untrained men.

This study compared the physiological strain induced by prolonged walking and running performed at the walk-run transition speed (WRTS) in healthy untrained men. Twenty volunteers (age: 28 ± 5.01 years; height: 174.0 ± 0.3 cm; body mass: 74.5 ± 0.6 kg) underwent the following: (a) ramp-incremental maximal cardiopulmonary exercise test (CPET); (b) specific protocol to detect the WRTS; and (c) two 30-min walking and running bouts at WRTS (mean ± SD: 6.9 ± 0.06 km/h). Expired gases were collected during exercise bouts via the metabolic cart. A significant effect of locomotion mode (F = 4.8, P < 0.001) was observed with running resulting in higher cardiorespiratory responses than walking at the WRTS (oxygen uptake: mean difference = 0.26 L/min; pulmonary ventilation: mean difference = 5.53 L/min; carbon dioxide output: mean difference = 0.32 L/min; heart rate: mean difference = 13 beats/min; total energy expenditure: mean difference = 59 kcal). The rating of perceived exertion was similar across locomotion modes (mean difference = 0.3; P = 0.490). In conclusion, running promoted greater cardiorespiratory responses than walking at the WRTS in untrained healthy men. These data might have practical impact on aerobic training performed at intensities corresponding to WRTS.

PERK inhibits DNA replication during the Unfolded Protein Response via Claspin and Chk1.

Stresses such as hypoxia, nutrient deprivation and acidification disturb protein folding in the endoplasmic reticulum (ER) and activate the Unfolded Protein Response (UPR) to trigger adaptive responses through the effectors, PERK, IRE1 and ATF6. Most of these responses relate to ER homoeostasis; however, here we show that the PERK branch of the UPR also controls DNA replication. Treatment of cells with the non-genotoxic UPR agonist thapsigargin led to a rapid inhibition of DNA synthesis that was attributable to a combination of DNA replication fork slowing and reduced replication origin firing. DNA synthesis inhibition was dependent on the UPR effector PERK and was associated with phosphorylation of the checkpoint adaptor protein Claspin and activation of the Chk1 effector kinase, both of which occurred in the absence of detectable DNA damage. Remarkably, thapsigargin did not inhibit bulk DNA synthesis or activate Chk1 in cells depleted of Claspin, or when Chk1 was depleted or subject to chemical inhibition. In each case thapsigargin-resistant DNA synthesis was due to an increase in replication origin firing that compensated for reduced fork progression. Taken together, our results unveil a new aspect of PERK function and previously unknown roles for Claspin and Chk1 as negative regulators of DNA replication in the absence of genotoxic stress. Because tumour cells proliferate in suboptimal environments, and frequently show evidence of UPR activation, this pathway could modulate the response to DNA replication-targeted chemotherapies.

Diferentes fontes de ferro na prevenção da anemia ferropriva e no desempenho de leitões lactentes / Different sources of iron for the prevention of iron deficiency anemia and performance of suckling piglets

Com o objetivo de avaliar o uso de diferentes fontes de ferro na prevenção da anemia ferropriva e no desempenho em leitões lactentes, dividiram-se 202 leitões em cinco tratamentos: FD - aplicação intramuscular de 200mg de ferro dextrano no terceiro dia de idade; T24 - terra à vontade fornecida aos leitões a cada 24 horas do terceiro ao 19º dia; T48 - terra à vontade fornecida aos leitões a cada 24 horas do terceiro ao 10º dia e do 11º ao 19º dia, com intervalo de 48 horas; T72 - terra à vontade fornecida aos leitões a cada 24 horas do terceiro ao 10º dia e do 11º ao 19º dia, com intervalo de 72 horas; SA - suplemento alimentar ultraprecoce rico em ferro quelatado em pó (SAUP) fornecido do terceiro ao 11º dia, com intervalo de 48 horas. O ferro dextrano aplicado no terceiro dia de vida e a suplementação com terra e SAUP foram eficientes para garantir o desempenho de leitões no período de aleitamento e não influenciaram no consumo de ração nem na taxa de viabilidade. As diferentes fontes de ferro estudadas não influenciaram o leucograma e foram eficientes na prevenção da anemia ferropriva e no desempenho dos leitões lactentes. Com relação às concentrações de hemoglobina e hematócrito, os animais suplementados com ferro dextrano apresentaram valores superiores quando comparados aos que recebem terra e SAUP.(AU)

In order to evaluate the use of different sources of iron to prevent iron deficiency anemia and to appraise the performance of suckling piglets, we sorted 202 piglets in five treatments. ID - intramuscular injection of 200mg of iron dextran on the third day of age; T24 - free daily access to land provided to piglets every 24 hours from the third to the nineteenth day; T48 - free daily access to land provided to piglets every 24 hours from the third to the tenth day and from day 11 to day 19 with an interval of 48 hours; T72 - free daily access to land provided to piglets every 24 hours from the third to the tenth day and from day 11 to day 19 with an interval of 72 hours; FS - Food supplement rich in iron-chelating powder (SAUP) available from the third to the eleventh day with an interval of 48 hours. The iron dextran applied on the third day of life as well as the supplementation with land and SAUP were effective to ensure the performance of piglets during the lactation period and did not affect feed intake or the viability rate. The different sources of iron studied did not influence the WBC (White Blood Cell) and succeded in preventing iron deficiency anemia and performance of suckling piglets. Regarding the concentrations of hemoglobin and hematocrit, the animals supplemented with iron dextran showed higher values when compared to those who receive land and SAUP.(AU)

RNF168 cooperates with RNF8 to mediate FOXM1 ubiquitination and degradation in breast cancer epirubicin treatment.

The forkhead box M1 (FOXM1) transcription factor has a central role in genotoxic agent response in breast cancer. FOXM1 is regulated at the post-translational level upon DNA damage, but the key mechanism involved remained enigmatic. RNF168 is a ubiquitination E3-ligase involved in DNA damage response. Western blot and gene promoter-reporter analyses showed that the expression level and transcriptional activity of FOXM1 reduced upon RNF168 overexpression and increased with RNF168 depletion by siRNA, suggesting that RNF168 negatively regulates FOXM1 expression. Co-immunoprecipitation studies in MCF-7 cells revealed that RNF168 interacted with FOXM1 and that upon epirubicin treatment FOXM1 downregulation was associated with an increase in RNF168 binding and conjugation to the protein degradation-associated K48-linked polyubiquitin chains. Consistently, RNF168 overexpression resulted in an increase in turnover of FOXM1 in MCF-7 cells treated with the protein synthesis inhibitor cycloheximide. Conversely, RNF168, knockdown significantly enhanced the half-life of FOXM1 in both absence and presence of epirubicin. Using a SUMOylation-defective FOXM1-5x(K>R) mutant, we demonstrated that SUMOylation is required for the recruitment of RNF168 to mediate FOXM1 degradation. In addition, clonogenic assays also showed that RNF168 mediates epirubicin action through targeting FOXM1, as RNF168 could synergise with epirubicin to repress clonal formation in wild-type but not in FOXM1-deficient mouse embryo fibroblasts (MEFs). The physiological relevance of RNF168-mediated FOXM1 repression is further emphasized by the significant inverse correlation between FOXM1 and RNF168 expression in breast cancer patient samples. Moreover, we also obtained evidence that RNF8 recruits RNF168 to FOXM1 upon epirubicin treatment and cooperates with RNF168 to catalyse FOXM1 ubiquitination and degradation. Collectively, these data suggest that RNF168 cooperates with RNF8 to mediate the ubiquitination and degradation of SUMOylated FOXM1 in breast cancer genotoxic response.

New insight on the underdrawing of 16th Flemish-Portuguese easel paintings by combined surface analysis and microanalytical techniques.

This paper focusses on the study of the underdrawings of 16th century easel paintings attributed to the workshop of the Portuguese-Flemish Master Frei Carlos. This investigation encompasses multidisciplinary research that relates the results of surface exams (infrared reflectography, standard light photography and infrared photography) with analytical investigations. The surface analysis of Frei Carlos' underdrawings by infrared reflectography has shown heterogeneous work, revealing two different situations: (1) an abundant and expressive underdrawing, revealing a Flemish influence and (2) a simple and outlined underdrawing. This preliminary research raised an important question related to this Portuguese-Flemish workshop and to the analytical approach: Is the underdrawing's heterogeneity, as observed in the reflectograms, related to different artists or is this rather an effect that is produced due to the use of different materials in the underdrawing's execution? Consequently, if different materials were used, how can we have access to the hidden underdrawings? In order to understand the reasons for this dissemblance, chemical analysis of micro-samples collected in underdrawing areas and representing both situations were carried out by optical microscopy, micro Fourier transform infrared spectroscopy (µ-FTIR), scanning electron microscopy coupled with energy dispersive X-ray spectrometry (SEM-EDX) and micro-Raman spectroscopy (µ-Raman). Taking into account the different possibilities and practical and theoretical limitations of surface and punctual examinations in the study of easel painting underdrawings, the methodology of research was adjusted, sometimes resulting in a re-analysis of experimental results. This research shows the importance of combining multispectral surface exams and chemical analysis in the understanding of the artistic creative processes of 16th century easel paintings.

OTUB1 inhibits the ubiquitination and degradation of FOXM1 in breast cancer and epirubicin resistance.

The forkhead transcription factor FOXM1 has a key role in DNA damage response, and its deregulated overexpression is associated with genotoxic drug resistance in breast cancer. However, little is known about the posttranslational mechanisms by which FOXM1 expression is regulated by genotoxic agents and how they are deregulated in resistant cells. Initial co-immunoprecipitation studies verified previous proteomic analysis finding that the OTUB1 is a novel FOXM1-interacting protein. Western blot analysis showed that both OTUB1 and FOXM1 expression reduced upon genotoxic agent treatment in MCF-7 cells, but remained relatively constant in resistant cells. FOXM1 expression reduced upon OTUB1 depletion by siRNA and increased with OTUB1 overexpression in MCF-7 cells, arguing that OTUB1 positively regulates FOXM1 expression. In agreement, co-immunoprecipitation experiments demonstrated that FOXM1 expression is associated with OTUB1 binding but inversely correlates with conjugation to the protein degradation-associated Lys-48-linked ubiquitin-chains. Overexpression of wild-type (WT) OTUB1, but not the OTUB1(C91S) mutant, disrupted the formation of Lys48-linked ubiquitin-conjugates on FOXM1. Importantly, knockdown of OTUB1 by siRNA resulted in an increase in turnover of FOXM1 in MCF-7 cells treated with the protein synthesis inhibitor cycloheximide, whereas overexpression of WT OTUB1, but not the OTUB1(C91S) mutant, significantly enhances the half-life of FOXM1. In addition, proliferative and clonogenic assays also show that OTUB1 can enhance the proliferative rate and epirubicin resistance through targeting FOXM1, as OTUB1 has little effect on FOXM1-deficient cells. The physiological relevance of the regulation of FOXM1 by OTUB1 is further underscored by the significant correlations between FOXM1 and OTUB1 expression in breast cancer patient samples. Cox-regression survival analysis indicates that OTUB1 overexpression is linked to poorer outcome in particular in patients treated with chemotherapy. Collectively, these data suggest that OTUB1 limits the ubiquitination and degradation of FOXM1 in breast cancer and has a key role in genotoxic agent resistance.

Wheat gluten intake increases weight gain and adiposity associated with reduced thermogenesis and energy expenditure in an animal model of obesity.

BACKGROUND/OBJECTIVES: The association between gluten and body weight is inconsistent. Previously, we showed that a gluten-free diet reduces weight gain without changing food intake in mice fed high-fat diets. In the present study, we investigated the effects of gluten intake on fat metabolism, thermogenesis and energy expenditure in mice fed a standard or high-fat diet. METHODS: Mice were fed four different experimental diets during 8 weeks: a control-standard diet (CD), a CD added with 4.5% of wheat gluten (CD-G), a high-fat diet (HFD) and a HFD added with 4.5% of wheat gluten (HFD-G). After 8 weeks, the mice received (99m)Tc-radiolabeled gluten orally to study gluten absorption and biodistribution or they underwent indirect calorimetry. After killing, subcutaneous and brown adipose tissues (SAT and BAT) were collected to assess thermogenesis-related protein expression. Lipid metabolism was studied in adipocyte cultures from the four groups. RESULTS: Despite having had the same energy intake, CD-G and HFD-G mice exhibited increased body weight and fat deposits compared with their respective controls. (99m)Tc-GLU or its peptides were detected in the blood, liver and visceral adipose tissue, suggesting that gluten can even reach extraintestinal organs. Uncoupling protein-1 expression was reduced in the BAT of HFD-G and in the SAT of CD-G and HFD-G mice. Indirect calorimetry showed lower oxygen volume consumption in CD-G and HFD-G groups compared with their controls. In HFD mice, daily energy expenditure was reduced with gluten intake. Gluten also reduced adiponectin, peroxisome proliferator-activated receptor (PPAR)-α and PPARγ and hormone-sensitive lipase in cultures of isolated adipocytes from HFD mice, whereas in the CD-G group, gluten intake increased interleukin-6 expression and tended to increase that of tumor necrosis factor. CONCLUSIONS: Wheat gluten promotes weight gain in animals on both HFD and CD, partly by reducing the thermogenic capacity of adipose tissues.

Aflatoxins ingestion and canine mammary tumors: There is an association?

The aim of this study was to determine the presence of mycotoxins on dogs feed and to explore the potential association between mycotoxins exposure and the chance of mamary tumors in a case-control study. The study included 256 female dogs from a hospital population, 85 with mammary tumors (case group) and 171 without mammary tumors (control group). An epidemiological questionnaire was applied to both groups, and the data were analyzed by the EpiInfo statistical package. For the study, 168 samples of the feed offered to dogs were analyzed for the presence of aflatoxins, fumonisins and zearalenone by high-performance liquid chromatography. Mycotoxins were found in 79 samples (100%) in the case group and 87/89 (97.8%) in the control group. Mycotoxins were detected in all types of feed, regardless feed quality. Level of aflatoxin B1 (p = 0.0356, OR = 2.74, 95%, CI 1.13 to 6.60), aflatoxin G1 (AFG1) (p = 0.00007, OR = 4.60, 95%, CI = 2.16 to 9.79), and aflatoxin G2 (AFG2) (p = 0.0133, OR = 9.91, 95%, CI 1.21 to 81.15) were statistically higher in case of mammary cancer. In contrast, neutering was a protective factor for mammary cancer (p = 0.0004, OR = 0.32, 95%, CI = 0.17 to 0.60).

Bicritical universality of the anisotropic Heisenberg model in a crystal field.

The bicritical properties of the three-dimensional classical anisotropic Heisenberg model in a crystal field are investigated through extensive Monte Carlo simulations on a simple cubic lattice, using Metropolis and Wolff algorithms. Field-mixing and multidimensional histogram techniques were employed in order to compute the probability distribution function of the extensive conjugate variables of interest and, using finite-size scaling analysis, the first-order transition line of the model was precisely located. The fourth-order cumulant of the order parameter was then calculated along this line and the bicritical point located with good precision from the cumulant crossings. The bicritical properties of this point were further investigated through the measurement of the universal probability distribution function of the order parameter. The results lead us to conclude that the studied bicritical point belongs in fact to the three-dimensional Heisenberg universality class.

USP29 controls the stability of checkpoint adaptor Claspin by deubiquitination.

The DNA damage checkpoint is essential for the maintenance of genome integrity after genotoxic stress, and also for cell survival in eukaryotes. Claspin has a key role in the ATR (ATM and Rad3-related)-Chk1 branch of the DNA damage checkpoint and is also required for correct DNA replication. To achieve properly these functions, Claspin is tightly regulated by ubiquitinin-dependent proteasomal degradation, which controls Claspin levels in a DNA-damage- and cell-cycle-dependent manner. Here, we identified a new regulator of Claspin, the ubiquitin-specific peptidase 29, USP29. Downregulation of USP29 destabilizes Claspin, whereas its overexpression promotes an increase in Claspin levels. USP29 interacts with Claspin and is able to deubiquitinate it both in vivo and in vitro. Most importantly, USP29 knockdown results in an impaired phosphorylation of Chk1 after DNA damage and USP29-depleted cells show a major defect in the S-phase progression. With these results, we identified USP29 as a new player in the ATR-Chk1 pathway and the control of DNA replication.