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Activity-based protein profiling (ABPP) is a versatile strategy for identifying and characterizing functional protein sites and compounds for therapeutic development. However, the vast majority of ABPP methods for covalent drug discovery target highly nucleophilic amino acids such as cysteine or lysine. Here, we report a methionine-directed ABPP platform using Redox-Activated Chemical Tagging (ReACT), which leverages a biomimetic oxidative ligation strategy for selective methionine modification. Application of ReACT to oncoprotein cyclin-dependent kinase 4 (CDK4) as a representative high-value drug target identified three new ligandable methionine sites. We then synthesized a methionine-targeting covalent ligand library bearing a diverse array of heterocyclic, heteroatom, and stereochemically rich substituents. ABPP screening of this focused library identified 1oxF11 as a covalent modifier of CDK4 at an allosteric M169 site. This compound inhibited kinase activity in a dose-dependent manner on purified protein and in breast cancer cells. Further investigation of 1oxF11 found prominent cation-π and H-bonding interactions stabilizing the binding of this fragment at the M169 site. Quantitative mass-spectrometry studies validated 1oxF11 ligation of CDK4 in breast cancer cell lysates. Further biochemical analyses revealed cross-talk between M169 oxidation and T172 phosphorylation, where M169 oxidation prevented phosphorylation of the activating T172 site on CDK4 and blocked cell cycle progression. By identifying a new mechanism for allosteric methionine redox regulation on CDK4 and developing a unique modality for its therapeutic intervention, this work showcases a generalizable platform that provides a starting point for engaging in broader chemoproteomics and protein ligand discovery efforts to find and target previously undruggable methionine sites.

Cell-cycle pathway impairments resulting in CDK4 and 6 activation are frequently observed in human papillomavirus (HPV)-negative squamous cell carcinoma of the head and neck (SCCHN). We investigated the activity of ribociclib, a CDK4/6 inhibitor, in SCCHN models with the aim of identifying predictive biomarkers of response. HPV-negative or HPV-positive SCCHN cell lines (n = 8) and patient-derived tumor xenograft (PDTX) models (n = 6) were used. The models were classified according to their sensitivity to ribociclib to investigate potential predictive biomarkers. Ribociclib had a cytostatic effect in some HPV-negative SCCHN models but had no effect in HPV-positive models. In SCCHN cell lines and PDTXs, the retinoblastoma (Rb) protein expression level correlated with ribociclib activity. Rb knockdown was, however, not sufficient to block G0–G1 arrest induced by ribociclib in Detroit-562 where p107, p130, and Forkhead BOX M1 (FOXM1) were also implicated in ribociclib activity. Cell lines harboring epithelial-to-mesenchymal transition (EMT) features were less sensitive to ribociclib than those with an epithelial phenotype. Rb downregulation induced EMT in our Rb-expressing SCCHN cell lines. However, ribociclib still had significant activity in one PDTX model with high Rb and vimentin expression, suggesting that the presence of vimentin alone is not enough to induce ribociclib resistance. These findings suggest that CDK4/6 inhibitors should be investigated in patients with HPV-negative SCCHN with high Rb expression and an epithelial phenotype. Although these biomarkers are not predictive in all cases, they may enrich the population that could benefit from CDK4/6 inhibitors.

Figure S2 shows the protein expression of Cyclin E1 in HPV-negative SCCHN human cell lines.

Supplementary Results, Figure 1 from Combined Inhibition of MEK and Mammalian Target of Rapamycin Abolishes Phosphorylation of Cyclin-Dependent Kinase 4 in Glioblastoma Cell Lines and Prevents Their Proliferation

Summary Christians often presume that immediate and universally extensive divine governance of human behavior is incompatible with human agency and responsibility. Against this presumption, Kathryn Tanner argues for a distinctive metalinguistic paradigm whereby Christians can coherently speak of God’s transcendence in such a way that divine action could never in principle ‘compete’ with human action. Thus, it is said, God can comprehensively will each human action without thereby compromising significant human freedom and corresponding moral responsibility. In this article, it is argued that Tanner’s non-competitivist paradigm fails to circumvent a theological version of the ‘direct argument’ for the incompatibility between human moral responsibility and causal determinism.

As long-period ground motion holds the key to understanding the interior of the Earth’s Moon, reducing long-period noise sources will be an essential area of focus in the design of future lunar seismometers. For the proposed Lunar Geophysical Network (LGN), the International Lunar Network (ILN) Science Definition Team specifies that an LGN enabling seismometer will need to be more sensitive than any previous seismometer at frequencies below 1 Hz. In an effort toward lowering the seismometer noise floor for lunar geophysical missions, we evaluate the 1/f Brownian noise and the temperature sensitivity of a seismometer. Temperature sensitivity of a seismometer is related to an important component of the seismometer output noise that is proportional to the temperature noise in the environment. The implications of the ILN requirement are presented in the context of the state-of-the-art InSight Seismic Experiment for Interior Structure (SEIS) Very Broad Band (VBB) planetary seismometer. Brownian noise due to internal friction was estimated for future lunar operation after accounting for the rebalance of the product of mass and distance to the center of gravity of the pendulum for the SEIS-VBB sensor. We find that Brownian noise could be a limiting factor in meeting the ILN requirement for lunar seismometers. Further, we have developed a formalism to understand the temperature sensitivity of a seismometer, relating it quantitatively to the local gravity, the thermoelastic coefficient of the spring, change in center of gravity, and the coefficient of thermal expansion of the mechanical structures. We found that in general the temperature sensitivity of a seismometer is proportional to the local gravity, and so the temperature sensitivity can be reduced when operating on a planetary body with lower gravity. Our Brownian noise and temperature sensitivity models will be useful in the design of the next generation of planetary seismometers.

Existing treatment strategies for pancreatobiliary malignancies are limited. Nowadays, surgery is the only path to cure these types of cancer, but only a small number of patients present with resectable tumors at the time of diagnosis. The notoriously poor prognosis, lack of diverse treatment options associated with pancreaticobiliary cancers, and their resistance to current therapies reflect the urge for the development of novel therapeutic targets. Cyclin-dependent kinase 4/6 (CDK4/6) inhibitors have emerged as an attractive therapeutic strategy in a number of cancers since their approval for treatment in patients with ER+/HER- breast cancer in combination with antiestrogens. In this article, we discuss the therapeutic potential of CDK4/6 inhibitors in pancreatobiliary cancers, notably cholangiocarcinoma and pancreatic ductal adenocarcinoma.

The classical genes are a member of immunoglobulin gene family that is involved in the presentation of antigen peptides. They are located in every nucleated cell surface, except in neurons and trophoblasts. They are synthesized in infected cell (viral infection), binding to viral proteins and giving rise to Swine Leucocyte Antigen (SLA) I- antigen complex. In the study, the SLA class I gene of Nigerian pigs with pigs from Kenya along with archived sequences from Asia and American pigs available on Genbank to gain a better understanding of matrilineal origin of Nigerian and Kenyan pigs. Gain insight into demographic distribution, their domestication and adaptation were compared. The deoxyribonucleic acid (DNA) was extracted from 90 pigs (60 Nigerian pigs (NP) and 30 pigs from Kenya) using Invitrogen Kit, quantified by using Nanodrop, and amplified by using Polymerase Chain Reaction. The expected bands were purified and sequenced for SLA class I. The sequences were aligned and phylogenetic tree was analyzed by using MEGA 6. BLAST (Basic Local Alignment Search Tools) was conducted in NCBI (National Center for Biotechnology Information). The sequences of Nigerian pigs and Kenya pigs were compared with published pig data in GenBank. Neighboring Joint (NJ) tree constructed with sequences from Nigeria, Kenya and other pigs from GenBank revealed that some of the NP clustered together with Asian pigs at the middle of the tree while some clustered just above the Kenyan below the tree and the outgroup homosapien stood out. The result from the calculation of total number of segregating sites gave the negative Tajima's D value (-0.181) while the non-significant difference (p > 0.05) obtained from the result showed that the null hypothesis was not rejected. In conclusion, there is genetic difference between the Nigerian and Kenyan pigs based on the phylogeny tree constructed. There is an excess of low frequency polymorphisms relative to population size. This may be as a result of evolutionary forces during course of adaptation in tropical countries in Africa.

Malignant pleural mesothelioma (MPM) is an aggressive cancer with limited therapeutic options. In this study, we evaluated the impact of CDK4/6 inhibition by palbociclib in a panel of 28 MPM cell lines, including 19 patient-derived cell lines, using a variety of approaches including RNA-sequencing. Palbociclib used alone sufficed to strongly and durably inhibit the proliferation of 23 MPM cell lines, indicating a unique sensitivity of MPM to CDK4/6 inhibition. Importantly, insensitivity to palbociclib was mostly explained by the lack of active T172-phosphorylated CDK4. This was associated with the high p16INK4A (CDKN2A) levels that accompany RB1 defects or inactivation, and also (unexpectedly) cyclin E1 over-expression in the presence of wild-type RB1. Prolonged treatment with palbociclib irreversibly inhibited proliferation despite re-induction of cell cycle genes upon drug washout. A senescence-associated secretory phenotype including various potentially immunogenic components was also irreversibly induced. Phosphorylated CDK4 was detected in 80% of 47 MPM tumors indicating their intrinsic sensitivity to CDK4/6 inhibitors. The absence of this phosphorylation in some highly proliferative MPM tumors was linked to partial deletions of RB1, leading to very high p16 (CDKN2A) expression. Our study strongly supports the clinical evaluation of CDK4/6 inhibitory drugs for MPM treatment, in monotherapy or combination therapy.

The success of shrimp farming lies in the proper monitoring of water quality in the pond. Of all the water quality parameters the dissolved oxygen plays a vital role in the proper growth of shrimps and influences mortality to a great extent. Traditional methods of sensing the water quality especially dissolved oxygen is done in laboratories far away from the actual pond site and it is often time consuming. Hence an accurate forecasting model which predicts the changes in the DO content is quintessential to decrease mortality, yield export quality shrimps and reduce operational costs. Water quality datasets are collected by using devices fabricated with wireless water quality sensors which floats in the ponds. Fuzzy C means was chosen as the clustering method based on the dataset collected and radial basis functions neural networks were constructed with accurate number of hidden neurons to predict the changes in the dissolved oxygen. Root Mean Square Error (RMSE), Mean Absolute Error (MAE), Correlation Coefficient (R) and Willmott index of agreement (WIA) were used as the evaluation parameters in comparison with the Multilayer Perceptron based Backpropagation Neural Network and Standard Radial Basis Function Neural Network for prediction of the DO content. The results show that the proposed method is effective with reduced errors and correlation coefficient close to unity. This accurate prediction of the dissolved oxygen helps the farmers to take corrective action when required and decrease the operational costs and produce export quality shrimps.

Cyclin-dependent kinase 4 (CDK4) is a master integrator that couples mitogenic/oncogenic signaling with the cell division cycle. It is deregulated in most cancers and inhibitors of CDK4 have become standard of care drugs for metastatic estrogen-receptor positive breast cancers and are being evaluated in a variety of other cancers. We previously characterized the T-loop phosphorylation at T172 of CDK4 as the highly regulated step that determines the activity of cyclin D-CDK4 complexes. Moreover we demonstrated that the highly variable detection of T172-phosphorylated CDK4 signals the presence or absence of the active CDK4 targeted by the CDK4/6 inhibitory drugs, which predicts the tumor cell sensitivity to these drugs including palbociclib. To date, the phosphorylation of CDK4 has been very poorly studied because only few biochemical techniques and reagents are available for it. In addition, the available ones including 2D-IEF separation of CDK4 modified forms are considered too tedious. The present report describes the generation, selection and characterization of the first monoclonal antibodies that specifically recognize the active CDK4 phosphorylated on its T172 residue. One key to this success was the immunization with a long phosphopeptide corresponding to the complete activation segment of CDK4. These monoclonal antibodies specifically recognize T172-phosphorylated CDK4 in a variety of assays, including western blotting, immunoprecipitation and, as a capture antibody, a sensitive ELISA from cell lysates. The specific immunoprecipitation of T172-phosphorylated CDK4 allowed to clarify the involvement of phosphorylations of co-immunoprecipitated p21 and p27, showing a privileged interaction of T172-phosphorylated CDK4 with S130-phosphorylated p21 and S10-phosphorylated p27.

Main text A key comparison has been made between the absorbed dose to water standards of the Physikalisch-Technische Bundesanstalt (PTB), Germany, and the BIPM, in accelerator photon beams. The results show the standards to be in agreement within the standard uncertainty of the comparison of 5.7 parts in 103. The results are analysed and presented in terms of degrees of equivalence, suitable for entry in the BIPM key comparison database. To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database https://www.bipm.org/kcdb/. The final report has been peer-reviewed and approved for publication by the CCRI, according to the provisions of the CIPM Mutual Recognition Arrangement (CIPM MRA).

Main text A first key comparison has been made between the air-kerma standards of the KRISS, Republic of Korea, and the BIPM in mammography x-ray beams. The results show the standards to be in agreement at the level of the expanded uncertainty for the comparison of 4.4 parts in 103. The results are analysed and presented in terms of degrees of equivalence, suitable for entry in the BIPM key comparison database. To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database https://www.bipm.org/kcdb/. The final report has been peer-reviewed and approved for publication by the CCRI, according to the provisions of the CIPM Mutual Recognition Arrangement (CIPM MRA).

Main text A key comparison has been made between the absorbed dose to water standards of the National Research Council of Canada (NRC), Canada and the BIPM in accelerator photon beams. The results show the standards to be in agreement within the standard uncertainty of the comparison of 5.8 parts in 103. The results are analysed and presented in terms of degrees of equivalence, suitable for entry in the BIPM key comparison database. To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database https://www.bipm.org/kcdb/. The final report has been peer-reviewed and approved for publication by the CCRI, according to the provisions of the CIPM Mutual Recognition Arrangement (CIPM MRA).

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Introduction Plusieurs examens histologiques extemporanes sur coupes congelees (EHEc) sont necessaires lors d’une chirurgie de Mohs. La microscopie confocale ex vivo en mode combine et coloration digitale (MCEVd) permet une visualisation anatomopathologique directe de prelevements entiers sans congelation ni coupe au cryostat. Cette technique parait plus rapide et moins contraignante que l’EHEc. Nous avons teste son interet dans l’analyse extemporanee de prelevements de carcinomes basocellulaires (CBCs) operes en chirurgie de Mohs. Materiel et methodes Cinq CBC de mauvais pronostic du visage ont ete inclus. Dix-sept galettes de Mohs ont ete successivement analysees en MCEVd (Vivascope 2500 4eme generation, Mavig, Allemagne) par des dermatologues recemment formes a la MCEVd puis en EHEc par des anatomopathologistes, pour rechercher la presence de foyers de CBC, identifier le sous-type histologique et caracteriser la topographie des foyers tumoraux residuels. La sensibilite et la specificite de la MCEVd etaient evaluees en comparaison de l’EHEc. Resultats L’EHEc detectait des foyers de CBC residuels dans 9 prelevements sur 17. La MCEVd les avait detectes sur 8 prelevements ce qui correspondait a une sensibilite de 89 % et une specificite de 88 %. A l’EHEc, 3/5 CBC etaient infiltrants et 2/5 nodulaires. En MCEVd, les CBCs etaient decrits comme nodulaires dans 4/5 cas et infiltrant dans 1/5 cas. La correlation topographique des foyers de CBC residuels entre les deux methodes d’analyse etait correcte dans 6 prelevements sur 8. Discussion La performance diagnostique de la MCEVd obtenue est superposable a celle rapportee dans la litterature avec d’autres techniques de microscopie confocale. L’acquisition de la technique etait facilitee par le mode combine reflectance/fluorescence et la recoloration digitale des images qui miment une coloration par hematoxyline et eosine d’un examen histologique classique. Les principales difficultes rencontrees ont ete d’obtenir un aplatissement correct des prelevements, notamment de l’epiderme et un bon ajustement des taux de fluorescence et reflectance lors de l’acquisition des images numeriques. Le faible score d’identification des sous-types histologiques s’explique en partie par notre courbe d’apprentissage. La topographique des foyers de CBC residuels differait rarement entre les deux methodes d’analyses alors que les prelevements examines en MCEVd sont entiers et non congeles contrairement a l’EHEc ou ils sont coupes apres fixation par congelation. Conclusion Notre etude, malgre son faible nombre de prelevements, montre la faisabilite de l’analyse des galettes de chirurgie de Mohs par MCEVd. Une etude de plus grande ampleur, incluant des CBCs agressifs notamment sclerodermiformes et infiltrants sera necessaire pour confirmer l’interet de la MCEVd dans cette indication.

Background Currently, metastatic colorectal cancer is treated as a homogeneous disease and only RAS mutational status has been approved as a negative predictive factor in patients treated with cetuximab. The aim of this study was to evaluate if recently identified molecular subtypes of colon cancer are associated with response of metastatic patients to first-line therapy. Patients and methods We collected and analysed 143 samples of human colorectal tumours with complete clinical annotations, including the response to treatment. Gene expression profiling was used to classify patients in three to six classes using four different molecular classifications. Correlations between molecular subtypes, response to treatment, progression-free and overall survival were analysed. Results We first demonstrated that the four previously described molecular classifications of colorectal cancer defined in non-metastatic patients also correctly classify stage IV patients. One of the classifications is strongly associated with response to FOLFIRI (P = 0.003), but not to FOLFOX (P = 0.911) and FOLFIRI + Bevacizumab (P = 0.190). In particular, we identify a molecular subtype representing 28% of the patients that shows an exceptionally high response rate to FOLFIRI (87.5%). These patients have a two-fold longer overall survival (40.1 months) when treated with FOLFIRI, as first-line regimen, instead of FOLFOX (18.6 months). Conclusions Our results demonstrate the interest of molecular classifications to develop tailored therapies for patients with metastatic colorectal cancer and a strong impact of the first-line regimen on the overall survival of some patients. This however remains to be confirmed in a large prospective clinical trial.

Main text A key comparison has been made between the absorbed dose to water standards of the Laboratoire National Henri Becquerel (LNE-LNHB), France and the BIPM in accelerator photon beams. The results show the standards to be in agreement within the standard uncertainty of the comparison of 5.9 parts in 103. The results are analysed and presented in terms of degrees of equivalence, suitable for entry in the BIPM key comparison database. To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database kcdb.bipm.org/. The final report has been peer-reviewed and approved for publication by the CCRI, according to the provisions of the CIPM Mutual Recognition Arrangement (CIPM MRA).

Introduction Il est recommande en France de faire une biopsie avant d’operer un carcinome basocellulaire (CBC) sauf si ce dernier est de diagnostic tres probable, de bon pronostic, et lorsque le geste chirurgical ne necessite pas de reconstruction complexe. La microscopie confocale ex vivo en mode combine (reflectance et fluorescence) et coloration digitale (MCEVd) permet aujourd’hui une analyse histologique immediate de biopsies sans fixation ni coupe. Nous avons evalue l’efficacite de la MCEVd a diagnostiquer, classer histologiquement les CBCs et determiner leur niveau d’invasion. Materiel et methodes Des punch-biopsies (3 mm) pre-operatoires de lesions suspectes de CBC ont ete prospectivement analysees de juillet a novembre 2019 en MCEVd (Vivascope 2500 4e generation, Mavig, Allemagne) par un interne de dermatologie forme a cette technique (O1) et par un dermatopathologiste (O2). L’examen de reference anatomopathologique traditionnel (gold standard -GS-) etait fait en aveugle de ces analyses. Le critere de jugement principal etait la confirmation du diagnostic de CBC. Les criteres de jugement secondaires etaient la classification en sous-types histologiques, le niveau d’invasion tumorale et le niveau de profondeur de la biopsie. La sensibilite (Se), la specificite (Sp) et les valeurs predictives positives (VPP) ou negatives (VPN) de la MCEVd ont ete determinees par rapport au GS. La concordance des diagnostics etait evaluee avec le coefficient Kappa de Cohen. Resultats Cent six punch-biopsies etaient realisees chez 66 patients dont 73 correspondaient a des CBCs avec le GS (17 superficiels, 33 nodulaires, 21 infiltrants et 1 sclerodermiforme). La MCEVd permettait une analyse en profondeur, par les deux observateurs, similaire au GS. Sur l’ensemble de ces prelevements, les Se et Sp de la MCEVd pour le diagnostic de CBC etaient respectivement pour l’O1 de 97 % et 86 % et pour l’O2 de 97 % et 94 % avec une VPP et VPN respectives de 96 % et 61 % pour l’O1 et 97 % et 91 % pour l’O2. La concordance d’identification du sous-type histologique etait bonne pour l’O1 (IC = 0,65 [0,54–0,76]) et l’O2 (IC = 0,78 [0,69–0,88]). La concordance d’evaluation de l’invasion tumorale etait moyenne pour l’O1 (IC = 0,58 [0,46–0,70]) et tres bonne pour l’O2 (IC = 0,81 [0,72–0,90]). Discussion Les performances diagnostiques de la MCEVd pour confirmer la presence de foyers tumoraux de CBC sont elevees. Son utilisation par l’anatomopathologiste est facilitee par la coloration digitale mimant la coloration H&E. La MCEVd permet d’analyser les punch-biopsies sur toute leur hauteur comme le GS et de bien apprecier le niveau d’invasion tumorale et le sous-type histologique. Conclusion Notre etude montre que la MCEVd permet une caracterisation extemporanee preoperatoire complete des CBCs ouvrant la voie a une prise en charge diagnostique et therapeutique acceleree de ces tumeurs.

Resume Le Reseau regional de reeducation et de readaptation pediatrique de la region Rhone-Alpes rassemble des professionnels impliques dans la prise en charge de l’enfant et de l’adolescent en situation de handicap. Ces professionnels sont au quotidien confrontes a des dilemmes qui mettent en jeu des decisions therapeutiques et les souhaits ou desirs de l’enfant et/ou de sa famille. Ainsi, lors du seminaire du reseau en 2011, deux situations cliniques presentees ont ete analysees par Catherine Perrotin a la lumiere de l’ethique. Les echanges qui s’en sont suivis ont mis en evidence l’interet de reflechir entre professionnels, soutenus par un intervenant, exterieur aux situations cliniques vecues sur le terrain. Afin de poursuivre cette reflexion, la commission ethique du reseau a organise des cafes ethiques. A ce jour, six cafes ethiques ont eu lieu a Lyon. Ils ont ete elabores a partir d’un theme choisi par la commission emanant de situations cliniques. Pour chaque reunion, la commission a sollicite un intervenant susceptible d’apporter un eclairage exterieur sur la problematique evoquee. Ces cafes sont devenus un espace de formation et de reflexion individuelle et collective permettant a tous les professionnels du reseau d’avoir des outils pour une demarche ethique dans leur pratique quotidienne.

In a recent paper, Seth Shabo sets out to show that logical renderings of the Direct Argument for incompatibilism about moral responsibility and causal determinism, an influential incompatibilist argument for this conclusion, fail. In particular, Shabo argues that the Direct Argument—cashed out in logical terms—fails because it rests on an invalid rule of inference, Rule B. Shabo argues that Rule B, rendered logically, is subject to a counterexample that he constructs. If he’s right about this, it follows that logical versions of the Direct Argument fail. In the paper, I argue that Shabo’s alleged counterexample to Rule B is unsuccessful. Since Shabo’s counterexample fails, I conclude that he has failed successfully to undermine logical versions of the Direct Argument.

A first key comparison has been made between the air-kerma standards of the NIM, Republic of China and the BIPM in mammography x-ray beams. The results show the standards to be in agreement at the level of the combined standard uncertainty of 2.8 parts in 103. The results are analysed and presented in terms of degrees of equivalence, suitable for entry in the BIPM key comparison database. Main text To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database kcdb.bipm.org/. The final report has been peer-reviewed and approved for publication by the CCRI, according to the provisions of the CIPM Mutual Recognition Arrangement (CIPM MRA).

A key comparison has been made between the absorbed dose to water standards of the KRISS, Korea and the BIPM in accelerator photon beams. The results show the standards to be in agreement within the expanded uncertainty (k = 2) of the comparison of 12 parts in 103. The results are analysed and presented in terms of degrees of equivalence, suitable for entry in the BIPM key comparison database. Main text To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database kcdb.bipm.org/. The final report has been peer-reviewed and approved for publication by the CCRI, according to the provisions of the CIPM Mutual Recognition Arrangement (CIPM MRA).

Although thyroid gland function is mainly under the control of pituitary TSH, other factors, such as iodine, play a role in this process. The thyroid is capable of producing different iodolipids such as 6-iodo-deltalactone and 2-iodohexadecanal (2-IHDA). It was shown that these iodolipids mimic some of the inhibitory effects of excess iodide on several thyroid parameters.To study the effect of 2-IHDA on cell proliferation and apoptosis in FRTL-5 cells.FRTL-5 cells were grown in the presence of TSH and treated with increasing concentrations of KI and 2-IHDA (0.5, 5, 10 and 33 µM) for 24, 48 and 72 h. Whereas KI inhibited cell proliferation only at 33 µM after 72 h of treatment, 2-IHDA inhibited in a time and concentration dependent manner. Analysis of cell cycle by flow cytometric DNA analysis revealed an accumulation of cells in G1 phase induced by 2-IHDA. The expression of cyclin A, cyclin D1 and cyclin D3 were reduced after treatment with 2-IHDA whereas CDK4 and CDK6 proteins were not modified. 2-IHDA induced a dynamic change in cytoplasmic to nuclear accumulation of p21 and p27 causing these proteins to be accumulated mostly in the nucleus. We also observed evidence of a pro-apoptotic effect of 2-IHDA at highest concentrations. No significant effect of KI was observed.These results suggest that the inhibitory effects of 2-IHDA on FRTL-5 thyroid cell proliferation are mediated by cell cycle arrest in G1/S phase and cell death by apoptosis.

Although TSH has been suggested to be a proliferative agent for thyrocytes, the effect of TSH on human thyroid cells remains controversial. In particular, most of the reported studies relied primarily on changes in DNA synthesis but have not included measurement of the number of cells. We argue that only a direct count of cell number, demonstrating classical exponential expansion, serves as a valid measurement of proliferation. Thus, although some data support TSH as a proliferative agent, most do not provide conclusive evidence. To generate conclusive evidence with regard to a proliferative effect of TSH in human thyrocytes, we performed various experiments using primary cultures of human thyrocytes. In contrast to previous reports, TSH [±insulin-like growth factor 1 (IGF-1)] did not induce proliferation of thyrocytes under a variety of different conditions. However, TSH/IGF-1 cotreatment did upregulate thyroid-specific gene expression including thyroglobulin (TG) and TSHR in a manner consistent with cellular differentiation. Evidence for a proliferative effect of TSH has been used to inform the American Thyroid Association’s guidelines for the management of thyroid cancer patients, which include TSH suppression. While these recommendations are admittedly based on low- to moderate-quality evidence, TSH suppression is still widely used. We present data that question the consensus view that TSH promotes proliferation of human thyrocytes (upon which the American Thyroid Association’s guidelines are based) and suggest that additional studies, including randomized controlled trials, are warranted to address this important clinical question.

The so-called Direct Argument for the incompatibility of moral responsibility and causal determinism depends on a rule of inference called Rule A, a rule that says no one is (or could be) even partly morally responsible for a necessary truth. While most philosophers think that Rule A is valid, Stephen Kearns has recently offered several alleged counterexamples to the rule. In the paper, I show that Kearns’ counterexamples are unsuccessful.

The BIPM graphite calorimeter standard for absorbed dose to water has been used in conjunction with an ionization chamber of known volume and with Monte Carlo simulations of these arrangements to determine the value for Wair in (60)Co radiation and in accelerator photon beams up to 25 MV. The results show no evidence for a variation in Wair at the 0.2% level over this energy range. Taking the constancy of Wair as established, the best estimate is Wair = 34.03 eV with a standard uncertainty of 0.21%. Consistent with this analysis, and assuming the use of the grain density in evaluating the stopping power of graphite, is the value Ic = 81.1 eV for the mean excitation energy for graphite, with standard uncertainty 1.8 eV.

A key comparison has been carried out of the absorbed dose to water standards of the Federal Institute of Metrology (METAS), Switzerland, and the BIPM in accelerator photon beams. The results show the standards to be in agreement within the standard uncertainty of the comparison of 6.6 parts in 103. The results are analysed and presented in terms of degrees of equivalence, suitable for entry in the BIPM key comparison database. KEY WORDS FOR SEARCH Ionizing radiation, dosimetry, key comparison, absorbed dose to water Main text To reach the main text of this paper, click on Final Report. Note that this text is that which appears in Appendix B of the BIPM key comparison database kcdb.bipm.org/. The final report has been peer-reviewed and approved for publication by the CCRI, according to the provisions of the CIPM Mutual Recognition Arrangement (CIPM MRA).

We present what is to our knowledge the first reported case of thoracic disc herniation leading to venous congestive myelopathy (VCM), which was clinically and radiographically suggestive of Foix-Alajouanine syndrome (angiodysgenetic necrotizing myelopathy). In addition, we review current concepts in evaluating the etiology of VCM and discuss indications for surgery.

The VIRGO Central Interferometer (CITF) is a short suspended interferometer operated with the central area elements of the VIRGO detector. The main motivation behind the CITF is to allow the integration and debugging of a large part of the subsystems of VIRGO while the construction of the long arms of the antenna is being completed. This will permit a faster commissioning of the full-size antenna. In fact, almost all the main components of the CITF, with the exception of the large mirrors and a few other details, are the same as those to be used for the full-size detector. In this paper the present status of the VIRGO CITF is reported.

Objective. To correlate long-term physical impairments of patients with severe traumatic brain injury (sTBI) based on their functional status in an acute care setting. Methods. 46 patients with sTBI participated in this prospective study. The Extended Glasgow Outcome Scale (GOSE) and the FIM instrument were rated at discharge from the acute care setting and at followup. The Functional Ambulation Classification (FAC), the Five-Meter Gait Speed, a quantified measure of negotiating stairs (Stair Climbing Speed and Rails used), and the functional reach test were rated at followup. Results. The subject with a score of 6 on the GOSE at discharge remained nonfunctional ambulator at followup. None of the subjects with a GOSE score of 5 became independent ambulators. Fifty percent of the subjects with a GOSE score of 4 were dependent ambulators. 100% of the subjects with a GOSE score of 2 or 3 at discharge were independent ambulators. A higher FIM score at discharge was associated with a greater chance of ambulating independently at 2 to 5 years after TBI (χKW22df). Conclusions. These data will allow physical health professionals in acute rehabilitation settings to provide more precise long-term physical outcome information to patients and families.

In his recent article, ‘Lottery puzzles and Jesus' return’, Donald Smith says that Christians should accept a very robust scepticism about the future because a Christian ought to think that the probability of Jesus' return happening at any future moment is inscrutable to her. But I think that Smith's argument lacks the power rationally to persuade Christians who are antecedently uncommitted as to whether or not we can or do have any substantive knowledge about the future. Moreover, I think that Christians who are so antecedently uncommitted have available objections they can reasonably press against Smith's arguments. In the article, I attempt to bring out these objections.

Although the specific CDK4/6 inhibitor PD0332991 (Palbociclib) was recently approved by the FDA to treat advanced ER+ breast tumors, there is yet no reliable sensitivity prediction tool. Cyclin D-CDK4/6 are the first CDK complexes to be activated in G1 phase in response to oncogenic pathways. They phosphorylate and inactivate the central cell cycle/tumor suppressor pRb. CDK4 activity requires its binding to a cyclin D (CCND1-3 genes) with which INK4 CDK4 inhibitors such as p16 (CDKN2A-D genes) compete. Although the assembly of the CDK4-cyclin D complexes was considered to be the main level of CDK4 activity control, we have shown that the activating T172-phosphorylation of CDK4 is actually the central rate-limiting event that initiates the cell cycle decision and signals the presence of active CDK4. Here, using 2D-gel electrophoresis to separate the modified forms of CDK4, we found in breast cancer cell lines that only the CDK4 T172-phosphorylation correlates with the sensitivity to PD0332991. The only exception was in the rare case of combined CCNE1 amplification and CDKN2A loss wherein combination of PD0332991 with a CDK2 inhibitor is required to block entry in the cell cycle. Additionally, three types of CDK4 modification profile were identified by 2D-gel electrophoresis in 56 breast tumors. In the first profile, the phosphorylated CDK4 was undetectable as in normal breast samples despite a high KI67 index. In the second and third profiles, the CDK4 phosphorylation was detectable and its intensity was either above or below 90% of the intensity of a second yet unidentified form of CDK4, respectively. The proportions of these profiles differ among breast tumors according to their clinic-pathological characteristics, molecular subtypes and risk. Finally, we identified a 11-gene expression signature that faithfully predicted the CDK4 modification profiles of breast tumors and cell lines (concordance rates of 84% and 100% in the 56 analyzed breast tumor samples or cell lines respectively). All three CDK4 modification profiles were evaluated in a merged independent dataset of 4034 published gene expression profiles. In these 4034 patients, 70% of triple-negative tumors, 18% of HER2-positive tumors and 5% of ER-positive tumors were predicted to have the first CDK4 profile wherein CDK4 phosphorylation is undetectable and to be completely unresponsive to CDK4 inhibitors. The phosphorylated CDK4 was predicted to be the major modified form in 26% of triple-negative tumors, 48% of HER2- positive tumors and 56% of ER-positive tumors. These patients should benefit the most from treatment with CDK4 inhibitors. Therefore, prediction of the CDK4 modification profile may allow extending treatment with Palbociclib to presently ineligible patients. As tumors with the third CDK4 modification profile generally present low grade and low OncotypeDX risks, the added value of including CDK4 inhibitors in their treatment compared to surgery and hormone therapy alone is questionable. In conclusion, we identified CDK4 phosphorylation as the most direct biomarker of CDK4 inhibitor sensitivity in breast cancer and developed a promising 11-gene based surrogate marker to guide their use in the clinic.Although the specific CDK4/6 inhibitor PD0332991 (Palbociclib) was recently approved by the FDA to treat advanced ER+ breast tumors, there is yet no reliable sensitivity prediction tool. Cyclin D-CDK4/6 are the first CDK complexes to be activated in G1 phase in response to oncogenic pathways. They phosphorylate and inactivate the central cell cycle/tumor suppressor pRb. CDK4 activity requires its binding to a cyclin D (CCND1-3 genes) with which INK4 CDK4 inhibitors such as p16 (CDKN2A-D genes) compete. Although the assembly of the CDK4-cyclin D complexes was considered to be the main level of CDK4 activity control, we have shown that the activating T172-phosphorylation of CDK4 is actually the central rate-limiting event that initiates the cell cycle decision and signals the presence of active CDK4. Here, using 2D-gel electrophoresis to separate the modified forms of CDK4, we found in breast cancer cell lines that only the CDK4 T172-phosphorylation correlates with the sensitivity to PD0332991. The only exception was in the rare case of combined CCNE1 amplification and CDKN2A loss wherein combination of PD0332991 with a CDK2 inhibitor is required to block entry in the cell cycle. Additionally, three types of CDK4 modification profile were identified by 2D-gel electrophoresis in 56 breast tumors. In the first profile, the phosphorylated CDK4 was undetectable as in normal breast samples despite a high KI67 index. In the second and third profiles, the CDK4 phosphorylation was detectable and its intensity was either above or below 90% of the intensity of a second yet unidentified form of CDK4, respectively. The proportions of these profiles differ among breast tumors according to their clinic-pathological characteristics, molecular subtypes and risk. Finally, we identified a 11-gene expression signature that faithfully predicted the CDK4 modification profiles of breast tumors and cell lines (concordance rates of 84% and 100% in the 56 analyzed breast tumor samples or cell lines respectively). All three CDK4 modification profiles were evaluated in a merged independent dataset of 4034 published gene expression profiles. In these 4034 patients, 70% of triple-negative tumors, 18% of HER2-positive tumors and 5% of ER-positive tumors were predicted to have the first CDK4 profile wherein CDK4 phosphorylation is undetectable and to be completely unresponsive to CDK4 inhibitors. The phosphorylated CDK4 was predicted to be the major modified form in 26% of triple-negative tumors, 48% of HER2- positive tumors and 56% of ER-positive tumors. These patients should benefit the most from treatment with CDK4 inhibitors. Therefore, prediction of the CDK4 modification profile may allow extending treatment with Palbociclib to presently ineligible patients. As tumors with the third CDK4 modification profile generally present low grade and low OncotypeDX risks, the added value of including CDK4 inhibitors in their treatment compared to surgery and hormone therapy alone is questionable. In conclusion, we identified CDK4 phosphorylation as the most direct biomarker of CDK4 inhibitor sensitivity in breast cancer and developed a promising 11-gene based surrogate marker to guide their use in the clinic. Citation Format: Raspe ES, Coulonval K, Pita J, Paternot S, Rothé F, Larsimont D, Van Laere S, Piccart M, Ignatiadis M, Sotiriou C, Roger PP. CDK4 phosphorylation status and corresponding gene expression profile predict sensitivity to Palbociclib [abstract]. In: Proceedings of the 2016 San Antonio Breast Cancer Symposium; 2016 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2017;77(4 Suppl):Abstract nr P6-07-02.

Mitosis is triggered by the abrupt dephosphorylation of inhibitory Y15 and T14 residues of cyclin B1-bound cyclin-dependent kinase (CDK)1 that is also phosphorylated at T161 in its activation loop. The sequence of events leading to the accumulation of fully phosphorylated cyclin B1-CDK1 complexes remains unclear. 2D-gel electrophoresis allowed us to discriminate whether T14, Y15 and T161 phosphorylations occur on same CDK1 molecules and to characterize the physiological occurrence of their seven phosphorylation combinations. Intriguingly, in cyclin B1-CDK1, the activating T161 phosphorylation never occurred without the T14 phosphorylation. This strict association could not be uncoupled by a substantial reduction of T14 phosphorylation in response to Myt1 knockdown, suggesting some causal relationship. However, T14 phosphorylation was not directly required for T161 phosphorylation, because Myt1 knockdown did uncouple them when cyclin B1-CDK1 complexes were prevented to accumulate in cytoplasm by leptomycin B. The coupling mechanism thus depended on unperturbed cyclin B1-CDK1 traffic. The unsuspected observation that the activating phosphorylation of cyclin B1-CDK1 was tightly coupled to its T14 phosphorylation, but not Y15 phosphorylation, suggests a mechanism protecting from premature activation by constitutively active CDK-activating kinase. This explained opposite impacts of reduced expression of Myt1 and Wee1 with only the latter inducing catastrophic mitoses., JOURNAL ARTICLE, SCOPUS: ar.j, info:eu-repo/semantics/published