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In this study, UiO-67 (Zr)/g-C 3 N 4 composites (U67N) were synthesized at wt.% ratios of 05:95, 15:85, and 30:70 using the solvothermal method at 80 °C for 24 h followed by calcination at 350 °C. The composites were characterized using UV-Vis diffuse reflectance spectroscopy, Fourier-transform infrared spectroscopy, photoluminescence spectroscopy, X-ray photoelectron spectroscopy, X-ray diffraction, scanning electron microscopy-energy-dispersive X-ray spectroscopy, transmission electron microscopy, and nitrogen physisorption analysis. In addition, thermal stability analysis of UiO-67 was conducted using thermogravimetric analysis. The photocatalytic performance of the composites was assessed during the degradation and mineralization of a mixture of methylparaben (MeP) and propylparaben (PrP) under simulated sunlight. The adsorption process of U67N 15:85 was characterized through kinetic studies and adsorption capacity experiments, which were modeled using pseudo-first-order and pseudo-second-order kinetics and Langmuir and Freundlich isotherms, respectively. The influence of pH levels 3, 5, and 7 on the photocatalytic degradation of the mixture was investigated, revealing enhanced degradation and mineralization at pH 3. The U67N composite exhibited dual capability in removing contaminants through adsorption and photocatalytic processes. Among the prepared composites, U67N 15:85 demonstrated the highest photocatalytic activity, achieving removal efficiencies of 96.8% for MeP, 92.5% for PrP, and 45.7% for total organic carbon in 300 kJ/m 2 accumulated energy (3 h of reaction time). The detoxification of the effluent was confirmed through acute toxicity evaluation using the Vibrio fischeri method. The oxidation mechanism of the heterojunction formed between UiO-67 (Zr) and g-C 3 N 4 was proposed based on PL analysis, photoelectrochemistry studies (including photocurrent response, Nyquist, and Mott-Schottky analyses), and scavenger assays., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

The electro-Fenton process (EFP) is a powerful advanced oxidation process beneficial to treating recalcitrant contaminants, and there has been a continuing interest in combining this technology to enhance the efficiency of conventional wastewater treatment processes. In this work, an optimized EFP process is performed as pretreatment for the degradation and mineralization of three blank fluoroquinolones (FQs) drugs: ofloxacin (OFL), norfloxacin (NOR), and ciprofloxacin (CIP). The optimization of the experiment was carried out using a Box-Behnken experimental design. Faster and complete degradation of the drugs mixture was achieved in 90 min with 61.12 ± 2.0% of mineralization in 180 min, under the optimized conditions: j = 244.0 mA cm -2 , [Fe 2+ ] = 0.31 mM, and [FQs] = 87.0 mg L -1 . Furthermore, a low toxicity effluent was obtained in 90 min of the experiment, according to bioassay toxicity with Vibrio fischeri. Five short-chain carboxylic acids, including oxalic, maleic, oxamic, formic, and fumaric acids, were detected and quantified, in addition to F - and NO 3 - inorganic ions. The inhibition of the reactive oxygen species with scavenger proof was also evaluated in this paper., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

The degradation of asulam herbicide by photo electro-Fenton (PEF) and solar photo electro-Fenton (SPEF) processes was studied using an undivided electrochemical BDD/carbon-felt cell to generate H 2 O 2 continuously. A central composite design combined with response surface methodology was applied to determine the optimal operating conditions of current intensity = 0.30 A, [Fe 2+ ] = 0.3 mM, and [Na 2 SO 4 ] = 0.11 M at pH 3 to achieve the complete degradation of asulam by electro-Fenton. Subsequently, the SPEF process was more efficient treatment compared to PEF, achieving a complete degradation of asulam and 98% of mineralization in 180 min. Moreover, 4-aminobenzenesulfonamide, 4-aminophenol, and 4-benzoquinone were detected as aromatic intermediates, whereas acetic acid, oxalic acid, and NO 3 - ions were identified as final degradation by-products. Thus, the SPEF process is an efficient alternative for the complete degradation and mineralization of herbicide asulam in an aqueous solution under natural sunlight., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier Ltd. All rights reserved.)

Aim: A series of knowledge has been developed on burnout syndrome in the sports context that has allowed to generate a solid theoretical structure that requires new contributions to delve into those aspects that have been less addressed, as is the case of optimism and resilience when it comes to linking them to the syndrome in top-performance athletes. For this purpose, the Burnout Inventory for Athletes (IBD-r), the Resilience Scale adapted to Spanish, and the Life Orientation Scale-Revised (LOT-R) for optimism were used., Methods: From this perspective, the study was approached with 121 Uruguayan athletes who participated in the last Pan-American Games, assuming practically all the subjects who have this level of sport (142) considering gender and type of sport., Results: The results indicate that 67% of the sample presents burnout symptoms for the emotional exhaustion dimension, and moderate resilience and optimism. Although there are no significant differences attributable to gender or type of sport, this contribution allows us to continue focusing on future work and further analysis. The type of sport presented statistically significant differences in relation to the personal competence dimension and the type of sport., Conclusion: Resilience and optimism obtained a clear influence on the occurrence of burnout, in a statistically negative sense, showing themselves as interesting prevention strategies for future lines of research, where it is essential to design interventions that teach emotional skills to manage adversity and prevent burnout., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Tutte-Vallarino, Malán-Ernst, Reyes-Bossio, Peinado-Portero, de Álvaro, Ortín Montero and Garcés de Los Fayos Ruiz.)

Excess weight requires multidisciplinary treatment, including psychology in face-to-face and e-health interventions. The need to analyse interventions' content validity has become evident. The objective is to assess content validity of a psychological e-health program of self-control and motivation for adults with excess weight. Six health professionals (a nutritionist, a personal trainer, physician and three psychologists) evaluated the content validity of the program. The results showed that the item-level content validity index (I-CVI) was 1 (the maximum value) for 11 of the 22 activities that comprise the program. This index fluctuated between 0.83 and 0.85 for the other activities, and the scale-level content validity index was greater than 0.90 for the four modules. In terms of relevance, effectiveness and appropriateness, 90%, 65% and 60% of the activities, respectively, obtained the maximum possible I-CVI, while the index was acceptable for the remainder of the activities. This is the first study about content validity of a psychological program for adults with excess weight in Mexico and it supported that the content program was suitable, and it may be used as a part of multidisciplinary treatments. This study also highlights the importance of involving an expert review in the development of intervention programs. This intervention program may be useful in the context of a multidisciplinary intervention; however, it is important to conduct studies to evaluate its efficacy., (© 2022 World Obesity Federation.)

Pancreatic ductal adenocarcinoma (PDAC) shows pronounced epithelial and mesenchymal cancer cell populations 1-4 . Cellular heterogeneity in PDAC is an important feature in disease subtype specification 3-5 , but how distinct PDAC subpopulations interact, and the molecular mechanisms that underlie PDAC cell fate decisions, are incompletely understood. Here we identify the BMP inhibitor GREM1 6,7 as a key regulator of cellular heterogeneity in pancreatic cancer in human and mouse. Grem1 inactivation in established PDAC in mice resulted in a direct conversion of epithelial into mesenchymal PDAC cells within days, suggesting that persistent GREM1 activity is required to maintain the epithelial PDAC subpopulations. By contrast, Grem1 overexpression caused an almost complete 'epithelialization' of highly mesenchymal PDAC, indicating that high GREM1 activity is sufficient to revert the mesenchymal fate of PDAC cells. Mechanistically, Grem1 was highly expressed in mesenchymal PDAC cells and inhibited the expression of the epithelial-mesenchymal transition transcription factors Snai1 (also known as Snail) and Snai2 (also known as Slug) in the epithelial cell compartment, therefore restricting epithelial-mesenchymal plasticity. Thus, constant suppression of BMP activity is essential to maintain epithelial PDAC cells, indicating that the maintenance of the cellular heterogeneity of pancreatic cancer requires continuous paracrine signalling elicited by a single soluble factor., (© 2022. The Author(s), under exclusive licence to Springer Nature Limited.)

Oxaliplatin is the first-line regime for advanced gastric cancer treatment, while its resistance is a major problem that leads to the failure of clinical treatments. Tumor cell heterogeneity has been considered as one of the main causes for drug resistance in cancer. In this study, the mechanism of oxaliplatin resistance was investigated through in vitro human gastric cancer organoids and gastric cancer oxaliplatin-resistant cell lines and in vivo subcutaneous tumorigenicity experiments. The in vitro and in vivo results indicated that CD133+ stem cell-like cells are the main subpopulation and PARP1 is the central gene mediating oxaliplatin resistance in gastric cancer. It was found that PARP1 can effectively repair DNA damage caused by oxaliplatin by means of mediating the opening of base excision repair pathway, leading to the occurrence of drug resistance. The CD133+ stem cells also exhibited upregulated expression of N6-methyladenosine (m6A) mRNA and its writer METTL3 as showed by immunoprecipitation followed by sequencing and transcriptome analysis. METTTL3 enhances the stability of PARP1 by recruiting YTHDF1 to target the 3'-untranslated Region (3'-UTR) of PARP1 mRNA. The CD133+ tumor stem cells can regulate the stability and expression of m6A to PARP1 through METTL3, and thus exerting the PARP1-mediated DNA damage repair ability. Therefore, our study demonstrated that m6A Methyltransferase METTL3 facilitates oxaliplatin resistance in CD133+ gastric cancer stem cells by Promoting PARP1 mRNA stability which increases base excision repair pathway activity., (© 2022. The Author(s), under exclusive licence to Springer Nature Switzerland AG.)

Lung squamous cell carcinoma (LSCC) is a considerable global health burden, with an incidence of over 600,000 cases per year. Treatment options are limited, and patient's 5-year survival rate is less than 5%. The ubiquitin-specific protease 28 (USP28) has been implicated in tumourigenesis through its stabilization of the oncoproteins c-MYC, c-JUN, and Δp63. Here, we show that genetic inactivation of Usp28 -induced regression of established murine LSCC lung tumours. We developed a small molecule that inhibits USP28 activity in the low nanomole range. While displaying cross-reactivity against the closest homologue USP25, this inhibitor showed a high degree of selectivity over other deubiquitinases. USP28 inhibitor treatment resulted in a dramatic decrease in c-MYC, c-JUN, and Δp63 proteins levels and consequently induced substantial regression of autochthonous murine LSCC tumours and human LSCC xenografts, thereby phenocopying the effect observed by genetic deletion. Thus, USP28 may represent a promising therapeutic target for the treatment of squamous cell lung carcinoma., Competing Interests: ER, AP, LL, TC, HS, AD, GV, ER, CD, NM, IR, EN, MG No competing interests declared, AT Andrew P Turnbull is affiliated with the CRUK Therapeutic Discovery Laboratories at the Crick Institute, for which no financial interests have been declared. APT declares competing financial interests due to financial support for the project described in this manuscript by Forma Therapeutics, Watertown, MA, USA. WK Wojciech W Krajewski is affiliated with the CRUK Therapeutic Discovery Laboratories at the Crick Institute, for which no financial interests have been declared. WWK declares competing financial interests due to financial support for the project described in this manuscript by Forma Therapeutics, Watertown, MA, USA. DG Dave Guerin is affiliated with Constellation Pharmaceuticals (Boston, USA), for which no financial interests have been declared. DG declares competing financial interests due to financial support for the project described in this manuscript by Forma Therapeutics, Watertown, MA, USA. JK Jeffrey Kearns is affiliated with the Novartis Institutes for BioMedical Research (Boston, USA), for which no financial interests have been declared. JK declares competing financial interests due to financial support for the project described in this manuscript by Forma Therapeutics, Watertown, MA, USA. SI Stephanos Ioannidis is affiliated with H3 Biomedicine (Cambridge, MA, USA), for which no financial interests have been declared. SI declares competing financial interests due to financial support for the project described in this manuscript by Forma Therapeutics, Watertown, MA, USA. MK Marie Katz is affiliated with Valo Health (Boston, USA), for which no financial interests have been declared. MK declares competing financial interests due to financial support for the project described in this manuscript by Forma Therapeutics, Watertown, MA, USA. CM Crystal McKinnon is affiliated with Valo Health (Boston, USA), for which no financial interests have been declared. CM declares competing financial interests due to financial support for the project described in this manuscript by Forma Therapeutics, Watertown, MA, USA. JO Johnathan O'Connell is affiliated with Valo Health (Boston, USA), for which no financial interests have been declared. JOC declares competing financial interests due to financial support for the project described in this manuscript by Forma Therapeutics, Watertown, MA, USA. NJ Neil Jones is affiliated with the CRUK Therapeutic Discovery Laboratories at the Crick Institute, for which no financial interests have been declared. NJ declares competing financial interests due to financial support for the project described in this manuscript by Forma Therapeutics, Watertown, MA, USA. CH Claire Heride is affiliated with the CRUK Therapeutic Discovery Laboratories at the Crick Institute, for which no financial interests have been declared. CH declares competing financial interests due to financial support for the project described in this manuscript by Forma Therapeutics, Watertown, MA, USA. MW Min Wu is affiliated with Disc Medicine (Cambridge, MA, USA), for which no financial interests have been declared. MW declares competing financial interests due to financial support for the project described in this manuscript by Forma Therapeutics, Watertown, MA, USA. CD Christopher J Dinsmore is affiliated with Disc Medicine (Cambridge, MA, USA), for which no financial interests have been declared. CJD declares competing financial interests due to financial support for the project described in this manuscript by Forma Therapeutics, Watertown, MA, USA. TH Tim R Hammonds is affiliated with the CRUK Therapeutic Discovery Laboratories at the Crick Institute, for which no financial interests have been declared. TRH declares competing financial interests due to financial support for the project described in this manuscript by Forma Therapeutics, Watertown, MA, USA. SK Sunkyu Kim is affiliated with Incyte (Wilmington, DE, USA), for which no financial interests have been declared. SK declares competing financial interests due to financial support for the project described in this manuscript by Forma Therapeutics, Watertown, MA, USA. DK DK declares competing financial interests due to financial support for the project described in this manuscript by Forma Therapeutics, Watertown, MA, USA. SU SU declares competing financial interests due to financial support for the project described in this manuscript by Forma Therapeutics, Watertown, MA, USA. MC MJC declares competing financial interests due to financial support for the project described in this manuscript by Forma Therapeutics, Watertown, MA, USA. BK BMK declares competing financial interests due to financial support for the project described in this manuscript by Forma Therapeutics, Watertown, MA, USA. AB AB declares competing financial interests due to financial support for the project described in this manuscript by Forma Therapeutics, Watertown, MA, USA., (© 2021, Ruiz et al.)

Oxaliplatin (OXA) resistance in the treatment of different types of cancer is an important and complex problem. The culture of tumor organoids derived from gastric cancer can help us to provide a deeper understanding of the underlying mechanisms that lead to OXA resistance. In this study, our purpose was to understand the mechanisms that lead to OXA resistance, and to provide survival benefits to patients with OXA through targeted combination therapies. Using sequence analysis of OXA-resistant and non-OXA-resistant organoids, we found that PARP1 is an important gene that mediates OXA resistance. Through the patients' follow-up data, it was observed that the expression level of PARP1 was significantly correlated with OXA resistance. This was confirmed by genetic manipulation of PARP1 expression in OXA-resistant organoids used in subcutaneous tumor formation. Results further showed that PARP1 mediated OXA resistance by inhibiting the base excision repair pathway. OXA also inhibited homologous recombination by CDK1 activity and importantly made cancers with normal BRCA1 function sensitive to PARP inhibition. As a result, combination of OXA and Olaparib (PARP-1/2/3 inhibitor), inhibited in vivo and in vitro OXA resistant organoid growth and viability., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Li, Wang, Lan, Wu, Evans, Ruiz, Yan, Zhou, Oliveira, Reis, Hu, Chen, Behrens, He and Zhang.)

The AP-1 transcription factor c-Jun is required for Ras-driven tumorigenesis in many tissues and is considered as a classical proto-oncogene. To determine the requirement for c-Jun in a mouse model of K-RasG12D-induced lung adenocarcinoma, we inducibly deleted c-Jun in the adult lung. Surprisingly, we found that inactivation of c-Jun, or mutation of its JNK phosphorylation sites, actually increased lung tumor burden. Mechanistically, we found that protein levels of the Jun family member JunD were increased in the absence of c-Jun. In c-Jun-deficient cells, JunD phosphorylation was increased, and expression of a dominant-active JNKK2-JNK1 transgene further increased lung tumor formation. Strikingly, deletion of JunD completely abolished Ras-driven lung tumorigenesis. This work identifies JunD, not c-Jun, as the crucial substrate of JNK signaling and oncogene required for Ras-induced lung cancer.

Cross-validation is a useful strategy to address problems associated with the post hoc model fitting. The aim of this study was to cross-validate the BAS-2, in two samples (Calibration and Validation Samples), using an invariance testing strategy. 1127 Mexican adolescents (508 males and 619 females), aged 11-19 years (M = 15.41, SD = 2.30), allocated in two samples (n 1  = 672; n 2  = 455) completed the Mexican version of the BAS-2, along with other body image-related instruments. The cross-validation supported the generalizability of the one-factor respecified model of the BAS-2. The measure was invariant across body mass index and age, but not across sex. Internal consistency and test-retest reliability (a three-week period between each test) was excellent. In support of the validity of the BAS-2, it was positively associated with scores on body esteem, but negatively with body mass index, drive for thinness, and body dissatisfaction. Normal-weight and young adolescents groups showed higher body appreciation than excess-weight and late adolescents groups, respectively. Considering these findings, it is reasonable to conclude that the BAS-2 is a reliable and valid instrument to assess body appreciation in Mexican adolescents. LEVEL OF EVIDENCE: Level V, cross-sectional, descriptive study.

Background: Stroke is an abrupt interruption of brain circulation that constitutes a neurological emergency. Aphasia is a language disorder caused by stroke. It can affect patients' reading, writing, comprehension and expression. The aim of the study is to find out if the use of the latest portable technologies such as tablets, computers and mobile phones can help to improve the rehabilitation of patients with aphasia., Methods: A literature review has been carried out on the main databases; Pubmed, Scopus and Cinahl, of articles in english and spanish published between june and december 2019 following the PRISMA methodology., Results: Sixty-two articles were included in which the use of technologies alongside traditional rehabilitation therapies showed that might improve the recovery of aphasic patients. Most studies have a sample size of less than fifty patients (21.7%), with only three publications having a sample size of more than 150 patients (1.86%)., Conclusions: The review shows that the use of digital applications might improve aphasia rehabilitation in patients who have suffered a stroke, but more studies are needed due to the small sample size of those conducted so far and to explore whether that improvement hold up over time.

Tumor cells proliferate rapidly and thus are frequently subjected to replication stress and the risk of incomplete duplication of the genome. Fragile sites are replicated late, making them more vulnerable to damage when DNA replication fails to complete. Therefore, genomic alterations at fragile sites are commonly observed in tumors. FRA16D is one of the most common fragile sites in lung cancer, however, the nature of the tumor suppressor genes affected by FRA16D alterations has been controversial. Here, we show that the ATMIN gene, which encodes a cofactor required for activation of ATM kinase by replication stress, is located close to FRA16D and is commonly lost in lung adenocarcinoma. Low ATMIN expression was frequently observed in human lung adenocarcinoma tumors and was associated with reduced patient survival, suggesting that ATMIN functions as a tumor suppressor in lung adenocarcinoma. Heterozygous Atmin deletion significantly increased tumor cell proliferation, tumor burden, and tumor grade in the LSL-KRasG12D; Trp53 F/F (KP) mouse model of lung adenocarcinoma, identifying ATMIN as a haploinsufficient tumor suppressor. ATMIN-deficient KP lung tumor cells showed increased survival in response to replication stress and consequently accumulated DNA damage. Thus, our data identify ATMIN as a key gene affected by genomic deletions at FRA16D in lung adenocarcinoma. SIGNIFICANCE: These findings identify ATMIN as a tumor suppressor in LUAD; fragility at chr16q23 correlates with loss of ATMIN in human LUAD and deletion of Atmin increases tumor burden in a LUAD mouse model., (©2019 American Association for Cancer Research.)

Sand swimming behaviour occurs in several lizard clades. Known ecological advantages of sand swimming include reduced predation risk and enhanced thermoregulation. We addressed whether, by way of sand abrasion, sand-swimming reduces ectoparasitism in the lizard Microlophus occipitalis, whose natural habitat includes sandy substrates (beach) and firm soil (dry forest). We hypothesised that, aside from habitat differences in infestation probability, ectoparasite prevalence and load would be lower in the beach than in the forest because of ectoparasite removal caused by sand-swimming. In an experiment with lizards confined in boxes with substrate from both habitats, lizards in beach boxes showed a greater decrease in ectoparasite load compared with lizards in forest boxes. Ectoparasite prevalence and load were much higher in the forest than in the beach across seasons. Larger lizards showed higher ectoparasite loads, and there were no sex differences in ectoparasite infestation. We provide evidence that sand swimming may confer another ecological advantage to lizards: reduced ectoparasitism.

Maternal separation (MS) early in life is related to an increase in anxiety and depressive-like behaviors and neurobiological alterations mostly related to alterations in hypothalamic pituitary adrenal (HPA) axis reactivity. Environmental enrichment (EE) has been used to ameliorate the effects of MS. However, the outcomes of this intervention at different developmental periods after MS have not been studied. We subjected male and female Sprague-Dawley pups to MS and subsequently compared the effects of EE started either in the pre-pubertal period [postnatal day (PND) 22] or adulthood (PND 78). Anxiety and depressive-like behaviors as well as in hippocampal synaptic density and basal corticosterone, oxytocin, and vasopressin levels were measured. Our results support the beneficial effects of adulthood EE in decreasing anxiety in males as well as promoting synaptic density in ventral hippocampal CA3. Males displayed higher levels of vasopressin while females displayed higher oxytocin, with no changes in basal corticosterone for any group after EE.

Non-steroidal anti-inflammatory drugs (NSAID) are compounds frequently found in municipal wastewater and their degradation by conventional wastewater treatment plants (WWTP) is generally incomplete. This study compared the efficiency of two advanced oxidation processes (AOP), namely heterogeneous photocatalysis (HP) and electro-Fenton (EF), in the degradation of a mixture of common NSAID (diclofenac, ibuprofen and naproxen) dissolved in either deionized water or effluent from a WWTP. Both processes were effective in degrading the NSAID mixture and the trend of degradation was as follows, diclofenac > naproxen > ibuprofen. EF with a current density of 40 mA cm -2 and 0.3 mmol Fe 2+  L -1 was the most efficient process to mineralize the organic compounds, achieving up to 92% TOC removal in deionized water and 90% in the WWTP effluent after 3 h of reaction. HP with 1.4 g TiO 2  L -1 at pH 7 under sunlight, produced 85% TOC removal in deionized water and 39% in WWTP effluent also after 3 h treatment. The lower TOC removal efficiency shown by HP with the WWTP effluent was attributed mainly to the scavenging of reactive species by background organic matter in the wastewater. On the contrary, inorganic ions in the wastewater may produce oxidazing species during the EF process, which contributes to a higher degradation efficiency. EF is a promising option for the treatment of anti-inflammatory pharmaceuticals in municipal WWTP at competitive electrical energy efficiencies.

Lung squamous cell carcinoma (LSCC) and adenocarcinoma (LADC) are the most common lung cancer subtypes. Molecular targeted treatments have improved LADC patient survival but are largely ineffective in LSCC. The tumor suppressor FBW7 is commonly mutated or down-regulated in human LSCC, and oncogenic KRasG12D activation combined with Fbxw7 inactivation in mice (KF model) caused both LSCC and LADC. Lineage-tracing experiments showed that CC10 + , but not basal, cells are the cells of origin of LSCC in KF mice. KF LSCC tumors recapitulated human LSCC resistance to cisplatin-based chemotherapy, and we identified LUBAC-mediated NF-κB signaling as a determinant of chemotherapy resistance in human and mouse. Inhibition of NF-κB activation using TAK1 or LUBAC inhibitors resensitized LSCC tumors to cisplatin, suggesting a future avenue for LSCC patient treatment., (© 2019 Ruiz et al.)

An automated method for the analysis of methylparaben, propylparaben, benzophenone-3, and benzophenone-4 in water effluents via on-line solid-phase extraction coupled with high-performance liquid chromatography/ultraviolet detection was proposed. The preconcentration parameters were studied using Plackett-Burman and Box-Behnken experimental designs using a C18 sorbent material. The results demonstrated that the eluent volume, composition, and sorbent amount were statistically significant. Optimal conditions for these variables were an eluent volume of 1.55 mL, eluent composition of acetonitrile 100% v/v, and sorbent amount of 100 mg. The eluted sample was analyzed on-line using high-performance liquid chromatography equipped with a reversed-phase C18 column and ultraviolet detection. Separation of the analytes was achieved in 15 min using gradient elution with acetonitrile/water. A simple, sensitive, and rapid analytical method was proposed for personal care compounds without sophisticated or expensive equipment. The limits of detection were 1.20, 1.73, 2.51, and 4.67 μg/L for propylparaben, methylparaben, benzophenone-3, and benzophenone-4, respectively. The analysis time was 48 min, consuming only 1.59 mL of eluent acetonitrile for the solid phase extraction step, with minimal sample handling. The method was applied to the analysis of spiked swimming pool and wastewater, with recoveries between 65-107%. These results indicate the reliability of the flow-based procedure., (© 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Purpose: The purpose of this study was to determine whether the muscle appearance satisfaction scale (MASS) shows acceptable psychometric properties in Mexican bodybuilders., Methods: A total of 258 Mexican male bodybuilders were recruited. Two self-report questionnaires, including the MASS and drive for muscularity scale (DMS), were administered. Six models of the latent structure of the MASS were evaluated, using confirmatory factor analysis with maximum likelihood, considering robust Satorra-Bentler correction to estimate the fit of the models to the data., Results: Similar to the original MASS, the series of CFA confirmed that the Mexican version was well represented with the 17-item five-factor structure, which showed a good model fit [Satorra-Bentler Chi-square (109, n = 258) = 189.18, p < 0.0001; NNFI = 0.91; CFI = 0.93; IFI = 0.93; RMSEA = 0.05 (0.04, 0.07)]. Internal consistency was estimated with McDonald's omega, which was acceptable for the MASS (0.88), and their subscales (0.80 to 0.89), except for muscle checking scale (0.77). Test-retest reliability analysis showed stability of the MASS total as well as of the subscale scores over a 2-week period (intraclass correlation coefficients = 0.75-0.91). Construct validity was demonstrated by a significant positive correlation between MASS and DMS results (r = 0.75; p = 0.0001). These results were similar to those of previous studies, which demonstrate the scale's usefulness., Conclusions: Our results support the suitability of the MASS and its subscales to measure muscle dysmorphia symptoms in Mexican male bodybuilders.

A commercial sulfamethoxazole + trimethoprim formulation has been degraded in 0.050 M Na 2 SO 4 at pH 3.0 by electrochemical oxidation with electrogenerated H 2 O 2 (EO-H 2 O 2 ), electro-Fenton (EF), photoelectro-Fenton with a 6-W UVA lamp (PEF) and solar photoelectro-Fenton (SPEF). The tests were performed in an undivided cell with an IrO 2 -based, Pt or boron-doped diamond (BDD) anode and an air-diffusion cathode for H 2 O 2 electrogeneration. The anode material had little effect on the accumulated H 2 O 2 concentration. Both drugs always obeyed a pseudo-first-order decay with low apparent rate constant in EO-H 2 O 2 . Much higher values were found in EF, PEF and SPEF, showing no difference because the main oxidant was always OH formed from Fenton's reaction between H 2 O 2 and added Fe 2+ . The solution mineralization increased in the sequence EO-H 2 O 2  < EF < PEF < SPEF regardless of the anode. The IrO 2 -based and Pt anodes behaved similarly but BDD was always more powerful. In SPEF, similar mineralization profiles were found for all anodes because of the rapid removal of photoactive intermediates by sunlight. About 87% mineralization was obtained as maximum for the powerful SPEF with BDD anode. Addition of Cl - enhanced the decay of both drugs due to their quicker reaction with generated active chlorine, but the formation of persistent chloroderivatives decelerated the mineralization process. Final carboxylic acids like oxalic and oxamic were detected, yielding Fe(III) complexes that remained stable in EF with BDD but were rapidly photolyzed in SPEF with BDD, explaining its superior mineralization ability., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

The present review documents the results of studies evaluating the acaricidal activity of different plant products and secondary metabolites against ticks that are resistant and susceptible to conventional acaricides. Studies published from 1998 to 2016 were included. The acaricidal activity of plant extracts, essential oils and secondary compounds from plants have been evaluated using bioassays with ticks in the larval and adult stages. There is variable effectiveness according to the species of plant and the concentrations used, with observed mortalities ranging from 5 to 100% against the Rhipicephalus (Boophilus), Amblyomma, Dermacentor, Hyalomma, and Argas genera. A number of plants have been reported to cause high mortalities and/or affect the reproductive capacity of ticks in the adult phase. In the majority of these trials, the main species of plants evaluated correspond to the families Lamiaceae, Fabaceae, Asteraceae, Piperaceae, Verbenaceae, and Poaceae. Different secondary metabolites such as thymol, carvacrol, 1,8-cineol and n-hexanal, have been found to be primarily responsible for the acaricidal activity of different essential oils against different species of ticks, while nicotine, dibenzyldisulfide and dibenzyltrisulfide have been evaluated for plant extracts. Only thymol, carvacrol and 1,8-cineol have been evaluated for acaricidal activity under in vivo conditions. The information in the present review allows the conclusion that the secondary metabolites contained in plant products could be used as an alternative for the control of ticks that are susceptible or resistant to commercial acaricides., (Copyright © 2017 Elsevier B.V. All rights reserved.)

This study reports the effect of putrescine addition, either alone or in combination with insulin, transferrin and selenite (ITS), to serum-free Advanced DMEM/F12 (A-DMEM/F12) medium, on the in vitro culture of Babesia bovis and using a perfusion bioreactor to improve efficiency of the process. A B. bovis strain previously adapted to proliferate in serum-free medium (Bbovis-SF) was evaluated using eight increasing concentrations of putrescine. The percentage of parasitized erythrocytes (PPE) obtained from cultures supplemented with 0.101 mg/L was 6.23% compared with 2.3% for control cultures with M199 with Earle's salts (M199) and 40% serum. The combination of putrescine (0.101 mg/L) and a mixture of ITS (2000, 1100, and 1.34 mg/L, respectively) (Pu-ITS), in A-DMEM/F12 culture medium without serum yielded a maximum PPE of 17.26% compared to 2.58% in the control medium. This new formulation of culture medium, together with the use of a hollow-fiber perfusion bioreactor system (HFPBS), caused a substantial increase in the proliferation of B. bovis, yielding a maximum cumulative PPE of 118.8% after five days, compared to 58.6% in cultures treated with control medium M199 and 40% serum. We concluded that the addition of the ITS mixture and putrescine to the culture medium stimulated the proliferation of B. bovis in vitro. This new medium formulation, used in a HFPBS culture system, can be an effective, automated-prone system that can induce massive proliferation of B. bovis for use as a source of parasite antigens and immunogens., (Copyright © 2017. Published by Elsevier Inc.)

Heritable thoracic aortic aneurysms and dissections (TAAD), including Marfan syndrome (MFS), currently lack a cure, and causative mutations have been identified for only a fraction of affected families. Here we identify the metalloproteinase ADAMTS1 and inducible nitric oxide synthase (NOS2) as therapeutic targets in individuals with TAAD. We show that Adamts1 is a major mediator of vascular homeostasis, given that genetic haploinsufficiency of Adamts1 in mice causes TAAD similar to MFS. Aortic nitric oxide and Nos2 levels were higher in Adamts1-deficient mice and in a mouse model of MFS (hereafter referred to as MFS mice), and Nos2 inactivation protected both types of mice from aortic pathology. Pharmacological inhibition of Nos2 rapidly reversed aortic dilation and medial degeneration in young Adamts1-deficient mice and in young or old MFS mice. Patients with MFS showed elevated NOS2 and decreased ADAMTS1 protein levels in the aorta. These findings uncover a possible causative role for the ADAMTS1-NOS2 axis in human TAAD and warrant evaluation of NOS2 inhibitors for therapy.

A 3.0 L solar flow plant with a Pt/air-diffusion (anode/cathode) cell, a solar photoreactor and a photocatalytic photoreactor filled with TiO2-coated glass spheres has been utilized to couple solar photoelectro-Fenton (SPEF) and solar heterogeneous photocatalysis (SPC) for treating a 165mgL(-1) salicylic acid solution of pH 3.0. Organics were destroyed by OH radicals formed on the TiO2 photocatalyst and at the Pt anode during water oxidation and in the bulk from Fenton's reaction between added Fe(2+) and cathodically generated H2O2, along with the photolytic action of sunlight. Poor salicylic acid removal and mineralization were attained using SPC, anodic oxidation with electrogenerated H2O2 (AO-H2O2) and coupled AO-H2O2-SPC. The electro-Fenton process accelerated the substrate decay, but with low mineralization by the formation of byproducts that are hardly destroyed by OH. The mineralization was strongly increased by SPEF due to the photolysis of products by sunlight, being enhanced by coupled SPEF-SPC due to the additional oxidation by OH at the TiO2 surface. The effect of current density on the performance of both processes was examined. The most potent SPEF-SPC process at 150mAcm(-2) yielded 87% mineralization and 13% current efficiency after consuming 6.0AhL(-1). Maleic, fumaric and oxalic acids detected as final carboxylic acids were completely removed by SPEF and SPEF-SPC., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Bovine serum is an important factor for the optimal growth of Babesia bovis in vitro. This protozoan can be cultured in M-199 with Earle's salts medium (M-199) supplemented with 40% bovine serum (BS). In the present study, four media were assessed along with the control medium M-199. The effect on the proliferation of B. bovis in vitro was tested when these media were combined with insulin (Ins), transferrin (Trans) and selenite (Sel) in the absence of bovine serum. Treatment with Advanced DMEM/F12 medium (A-DMEM/F12) achieved the highest percentage of parasitized erythrocytes (PPE), reaching a maximum value of 9.59%. A-DMEM/F12 medium supplemented with a mixture of Ins (2000 mg/L), Trans (1100 mg/L), and Sel (1.34 mg/L) allowed for the adaptation and proliferation of B. bovis without bovine serum, showed a constant increase in PPE, and reached a maximum value of 9.7% during seven cycles of in vitro culture. It was concluded that continuous proliferation of B. bovis in vitro could be achieved using A-DMEM/F12 medium supplemented with Ins-Trans-Sel, without bovine serum. After adaptation for proliferation in serum-free medium, the B. bovis strain of parasites could have future use in the study of this economically important protozoan species that affects cattle., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Acute exposure to morphine after a traumatic event reduces trauma related symptoms in humans and conditioned fear expression in male rats. We aimed to determine whether acute administration of morphine alters consolidation of fear learning and extinction. Male and female rats in proestrus and metaestrus (high and low ovarian hormones respectively) underwent fear conditioning and received saline or morphine (2.5 mg/kg s.c.). The next day they underwent extinction. Results showed increased freezing during extinction only in the morphine metaestrus group while morphine did not affect males or proestrus females. Recall of extinction was similar on all groups. On a second experiment, a subset of rats conditioned during metaestrus was administered morphine prior to extinction producing no effects. We then measured mu opioid receptor (MOR) expression in the amygdala and periaqueductal gray (PAG) at the end of extinction (day 2). In males and proestrus females, morphine caused an increase in MOR in the amygdala but no in the PAG. In metaestrus females, morphine did not change MOR expression in either structure. These data suggests that ovarian hormones may interact with MORs in the amygdala to transiently alter memory consolidation. Morphine given after trauma to females with low ovarian hormones might increase the recall of fear responses, making recovery harder.

Here, the synergetic effect of coupling solar photoelectro-Fenton (SPEF) and solar heterogeneous photocatalysis (SPC) on the mineralization of 200mL of a 20mg L(-1) atrazine solution, prepared from the commercial herbicide Gesaprim, at pH 3.0 was studied. Uniform, homogeneous and adherent anatase-TiO2 films onto glass spheres of 5mm diameter were prepared by the sol-gel dip-coating method and used as catalyst for SPC. However, this procedure yielded a poor removal of the substrate because of the low oxidation ability of positive holes and OH formed at the catalyst surface to destroy it. Atrazine decay was improved using anodic oxidation (AO), electro-Fenton (EF), SPEF and coupled SPEF-SPC at 100mA. The electrolytic cell contained a boron-doped diamond (BDD) anode and H2O2 was generated at a BDD cathode fed with an air flow. The removal and mineralization of atrazine increased when more oxidizing agents were generated in the sequence AO

The signals that regulate stem cell self-renewal and differentiation in the lung remain elusive. Lung stem cells undergo self-renewal or lineage commitment to replenish tissue, depending on cross-talk with their environment. This environment, also known as the niche, includes mesenchymal and endothelial tissues. Here we define molecular mechanisms involved in the interaction between human lung Lgr6+ stem cells (LSCs) and fibroblasts in a functional microenvironment. We reveal a central role for p38α MAPK in establishing and maintaining such cross-talk, acting in both cell types. In LSCs, p38α induces the expression of SDF-1, which activates the stroma. p38α is essential for fibroblast activation and induction of cytokine expression, in particular TNFα. This paracrine network induces a hierarchical activation leading to the recruitment of endothelium, establishing a functional microenvironment. Disruption of this cross-talk abrogates proper LSC differentiation in vivo and may lead to lung dysfunction and disease.

Background: Environmental stressors require the professional diver to undergo a complex process of psychophysiological adaptation in order to overcome the demands of an extreme environment and carry out effective and efficient work under water. The influence of cognitive and personality traits in predicting underwater performance and adaptation has been a common concern for diving psychology, and definitive conclusions have not been reached., Method: In this ex post facto study, psychological and academic data were analyzed from a large sample of personnel participating in scuba diving courses carried out in the Spanish Navy Diving Center. In order to verify the relevance of individual differences in adaptation to a hostile environment, we evaluated the predictive validity of general mental ability and personality traits with regression techniques., Results: The data indicated the existence of psychological variables that can predict the performance ( R² = .30, p <.001) and adaptation ( R²(N) = .51, p <.001) of divers in underwater environment., Conclusions: These findings support the hypothesis that individual differences are related to the probability of successful adaptation and effective performance in professional diving. These results also verify that dispositional traits play a decisive role in diving training and are significant factors in divers' psychological fitness.

One hundred and sixty 1-month-old turkey poults were delivered to 40 households in four communities of the State of Yucatan, Mexico. The poults were divided into two populations, one vaccinated and the other non-vaccinated against avian pox. During three months, monthly visits were carried out in order to monitor the appearance of lesions suggesting avian pox in the birds delivered. Each turkey was clinically examined, searching for characteristic avian pox lesions that were classified according to the degree of severity observed. The true incidence rate and the cumulative incidence rate of avian pox were determined and the true incidence and cumulative incidence rates of mortality were determined and the relative risks calculated. The true incidence rates for avian pox in vaccinated and non-vaccinated birds were 1.5 and 1.47 respectively. The cumulative incidence rates were 0.94 and 0.90 for vaccinated and non-vaccinated birds, respectively. The comparison for the whole period between vaccinated and non-vaccinated groups did not show a significant statistical difference for mortality. However, when mortality was compared between vaccinated and non-vaccinated turkeys for each month of the study, there was a statistically significant difference for the first month (relative risk = 0.216, confidence interval 0.069 to 0.676). In addition, when the severity of pox lesions between groups was compared, statistically significant differences were found in favour of the vaccinated birds (P < 0.0001).

Lung cancer is the cause of most cancer-related deaths in the Western world. Non-small cell lung cancer accounts for almost 80% of all lung cancers, and 50% of this type are adenocarcinomas. The cellular and molecular origin of this type of lung cancer remains elusive and the mechanisms are poorly known. It is known that K-Ras mutations appear in 25-30% of lung adenocarcinomas and it is the best known single mutation that can be related to lung cancers. Recently, it has been suggested that a putative population of mouse bronchioalveolar stem cells could be considered as the cell of origin of adenocarcinomas. These cells are expanded in the early stages of lung tumorigenesis. We have isolated a population of mouse bronchioalveolar stem cells and induced their transformation by oncogenic K-RasG12. Different approaches have shown that an intracellular network linking the p38α MAPK and the PI3K-Pdk1 pathways is involved in regulating the survival and malignant progression of the transformed cells. Absence of p38α catalytic activity leads to further Pdk1 activation (independent of Akt and Erk activity), enhancing the survival and proliferation of the more malignant lung cancer cells. This specifically selects high Sca-1/Sox9 cells that harbour a stronger colonizing potential, as they maintain their capacity to produce secondary tumors after serial transplantations.

This study refers to the evaluation of the psychometric properties of the Drive for Muscularity Scale among Mexican population. The exploratory factor analysis was carried out with 369 university males (mean age = 20.93 years). A three-factor structure that explains 45.64 % of the variance was found for the scale: 31.10 % by attitudes (α = 0.87), 10.70 % by substance intake (α = 0.72) and 3.84 % by training adherence (α = 0.68). Only the two first sub-scales and total score (α = 0.86) offered acceptable levels of internal consistence. The confirmatory factor analysis, carried out with 200 males (mean age = 20.79 years), verified both the suitability of this factor structure and the two-factor structure proposed by McCreary et al. (Psychol Men Masc 5:49-58, 1). Drive for muscularity is important in comprehending male body image and Drive for Muscularity Scale has demonstrated to be psychometrically suitable to assess this construct in Mexican males.

This article summarizes the current situation of one of the most important XXI century epidemics. The high rates of obesity in all countries and specially in our country at early ages are worrying. Traditionally, investigations have focused on the most nutritional aspects, such us dietary factors, metabolism, calory intake and energy expenditure. However, other sciences -such as Psychology- point out new influencing factors, such as food addiction, depression, stress and anxiety. In this piece of work, more than 80 articles from all disciplines and different databases and related to obesity are analyzed, as well as a new theoretical approach is proposed to ensure a more accurate comprehension. The literature review shows the necessary evidence to introduce a new model of obesity named edorexia, and raise issues to be resolved in future research oriented from a multidisciplinary perspective.

Traditionally phenology studies have been focused on changes through time, but there exist many instances in ecological research where it is necessary to interpolate among spatially stratified samples. The combined use of Geographical Information Systems (GIS) and Geostatistics can be an essential tool for spatial analysis in phenological studies. Geostatistics are a family of statistics that describe correlations through space/time and they can be used for both quantifying spatial correlation and interpolating unsampled points. In the present work, estimations based upon Geostatistics and GIS mapping have enabled the construction of spatial models that reflect phenological evolution of Vulpia geniculata (L.) Link throughout the study area during sampling season. Ten sampling points, scattered throughout the city and low mountains in the "Sierra de Córdoba" were chosen to carry out the weekly phenological monitoring during flowering season. The phenological data were interpolated by applying the traditional geostatitical method of Kriging, which was used to elaborate weekly estimations of V. geniculata phenology in unsampled areas. Finally, the application of Geostatistics and GIS to create phenological maps could be an essential complement in pollen aerobiological studies, given the increased interest in obtaining automatic aerobiological forecasting maps.

The rapidly destructive osteoarthritis of the hip is a variant of osteoarthritis that is characterized by hip pain during one to twelve months and rapid destruction of the femoral head, in absence of infectious, neurologic, metabolic or inflammatory disease. We present the case of a 66 year- old male with severe COPD that referred pain in both hips and was unable to walk. In the radiography of the pelvis complete destruction of both femoral heads was observed. The patient did not have abnormalities in blood tests that were related to infectious, neurologic or metabolic diseases. The patient underwent bilateral total hip replacement with prosthesis and physical therapy later. The patient functional status greatly improved with this treatment., (Copyright © 2009 Elsevier España, S.L. All rights reserved.)

The degradation of the Acid Yellow 36 (AY36) azo dye is studied by electro-Fenton (EF) and solar photoelectro-Fenton (SPEF) using a recirculation flow plant with an undivided cell containing a boron-doped diamond anode and an air-diffusion cathode for H₂O₂ electrogeneration, coupled with a solar photoreactor. A solution of 2.5L with 108 mg L⁻¹ of the dye and 0.5 mM Fe²(+) at pH 3.0 was comparatively treated at constant current. Hydroxyl radicals formed from Fenton's reaction and at the anode surface are the main oxidants. Total mineralization is almost achieved in SPEF, while EF yields poor TOC removal. Both processes are accelerated with increasing current. AY36 decays with similar rate in EF and SPEF following a pseudo first-order reaction, but the solution is more slowly decolorized because of the formation of conjugated byproducts. NH₄(+) ion is released in SPEF, while NO₃⁻ ion is mainly lost in EF. Tartronic, maleic, fumaric, oxalic, formic and oxamic acids are detected as generated carboxylic acids. Fe(III)-oxalate complexes are largely accumulated in EF and their quick photodecomposition in SPEF explains its higher oxidation power. The SPEF method yields greater current efficiency and lower energy cost as current decreases, and then it is more viable at low currents., (Copyright © 2010 Elsevier Ltd. All rights reserved.)

Activation of CDK1 is essential for M-phase entry both in mitosis and meiosis. G2-arrested oocytes contain a pool of CDK1/cyclin B complexes that are maintained inactive because of the phosphorylation of CDK1 on Thr14 and Tyr15 by the Wee1 family protein kinase Myt1, whose inhibition suffices to induce meiosis I entry [1-5]. CDK1/XRINGO and p90Rsk can both phosphorylate and downregulate Myt1 activity in vitro [6, 7]. Here we identify five p90Rsk phosphorylation sites on Myt1 that are different from the CDK1/XRINGO sites, and we show how both kinases synergize during oocyte maturation to inhibit Myt1, ensuring meiotic progression. We found that phosphorylation of Myt1 by CDK1/XRINGO early during oocyte maturation not only downregulates Myt1 kinase activity but also facilitates the recruitment of p90Rsk and further phosphorylation of Myt1. Mutation of the five p90Rsk residues to alanine impairs Myt1 hyperphosphorylation during oocyte maturation and makes Myt1 resistant to the inhibition by p90Rsk. Importantly, Myt1 phosphorylated by p90Rsk does not interact with CDK1/cyclin B, ensuring that the inhibitory phosphorylations of CDK1 cannot take place after meiosis I entry and contributing to the all-or-none meiotic response., (Copyright © 2010 Elsevier Ltd. All rights reserved.)

Cell-cycle progression is regulated by cyclin-dependent kinases (CDKs). CDK1 and CDK2 can be also activated by noncyclin proteins named RINGO/Speedy, which were identified as inducers of the G2/M transition in Xenopus oocytes. However, it is unclear how XRINGO triggers M phase entry in oocytes. We show here that XRINGO-activated CDKs can phosphorylate specific residues in the regulatory domain of Myt1, a Wee1 family kinase that plays a key role in the G2 arrest of oocytes. We have identified three Ser that are major phosphoacceptor sites for CDK/XRINGO but are poorly phosphorylated by CDK/cyclin. Phosphorylation of these Ser inhibits Myt1 activity, whereas their mutation makes Myt1 resistant to inhibition by CDK/XRINGO. Our results demonstrate that XRINGO-activated CDKs have different substrate specificity than the CDK/cyclin complexes. We also describe a mechanism of Myt1 regulation based on site-specific phosphorylation, which is likely to mediate the induction of G2/M transition in oocytes by XRINGO.

Identifying 14-3-3 isoform-specific substrates and functions may be of broad relevance to cell signaling research because of the key role played by this family of proteins in many vital processes. A multitude of ligands have been identified, but the extent to which they are isoform-specific is a matter of debate. Herein we demonstrate, both in vitro and in vivo, a specific, functionally relevant interaction of human 14-3-3gamma with the molecular scaffold KSR1, which is mediated by the C-terminal stretch of 14-3-3gamma. Specific binding to 14-3-3gamma protected KSR1 from epidermal growth factor-induced dephosphorylation and impaired its ability to activate ERK2 and facilitate Ras signaling in Xenopus oocytes. Furthermore, RNA interference-mediated inhibition of 14-3-3gamma resulted in the accumulation of KSR1 in the plasma membrane, all in accordance with 14-3-3gamma being the cytosolic anchor that keeps KSR1 inactive. We also provide evidence that KSR1-bound 14-3-3gamma heterodimerized preferentially with selected isoforms and that KSR1 bound monomeric 14-3-3gamma. In sum, we have demonstrated ligand discrimination among 14-3-3 isoforms and shed light on molecular mechanisms of 14-3-3 functional specificity and KSR1 regulation.

Cell-cycle transitions are controlled by CDKs (cyclin-dependent kinases), whose activation is usually associated with the binding of cyclins. RINGO/Speedy proteins can also bind to and activate CDKs, although they do not have amino acid sequence homology with cyclins. The RINGO/Speedy family members studied so far positively regulate cell-cycle progression. In the present paper, we report the biochemical and functional characterization of RINGO/Speedy E. We show that RINGO/Speedy E is a functionally distant member of this protein family that negatively affects cell-cycle progression. RINGO/Speedy E overexpression inhibits the meiotic progression in Xenopus oocytes as well as the proliferation of mammalian cells. RINGO/Speedy E can bind to endogenous CDK1 and CDK2 in both cellular systems. However, the RINGO/Speedy E-activated CDKs have different substrate specificity than the CDKs activated by other RINGO/Speedy proteins, which may account for their different effects on the cell cycle. Our results indicate that, although all RINGO/Speedy family members can activate CDKs, they may differently regulate cell-cycle progression.

We propose here a new strategy for the exhaustive mapping of phosphorylation sites in the Xenopus laevis Cdc25 phosphatase, which regulates cell cycle progression in eukaryotic cells. Two different MS analyses in a linear IT were used to identify the phosphorylated residues. First, a data-dependent neutral loss (DDNL) analysis triggered the fragmentation of peptides that show enhanced neutral loss of phosphoric acid. Second, a targeted product ion scanning (TPIS) mass analysis was carried out in which MS2 events are triggered for specific m/z values. Full coverage of the protein sequence was obtained by combining the two analyses with two enzymatic digestions, trypsin and chymotrypsin, yielding a comprehensive map of the phosphorylation sites. Previous reports have shown Cdc25C to be phosphorylated by Cdc2-cyclin B at four residues (Thr48, Thr67, Thr138 and Ser205). By using this combination of scan modes, we have identified four additional phosphorylation sites (Thr86, Ser99, Thr112 and Ser163) in a recombinant Cdc25C protein containing 198 residues of the NH2-terminal noncatalytic domain. The sensitivity of this combined approach makes it extremely useful for the comprehensive characterization of phosphorylation sites, virtually permitting complete coverage of the protein sequence with peptides within the mass detection range of the linear IT.

Cryptophane cages serve as host molecules to a Xe atom. Functionalization of cryptophane-A has permitted the development of Xe as a biosensor. Synthetic routes used to prepare cryptophanes result in racemic mixtures of the chiral cages. In the preparation of a tethered cryptophane-A cage for biosensor applications, some achiral and chiral substituents such as left-handed amino acids have been used. When the substituent is achiral, the NMR signal of the Xe atom in the functionalized cage in solution is a single isotropic peak, since the Xe shielding tensor components in the R and L cages differ by no more than the signs of the off-diagonal elements. Chiral substituents can split the cage-encapsulated Xe NMR signal into one or more sets of doublets, depending on the number of asymmetric centers in the substituent. We carry out quantum mechanical calculations of Xe nuclear magnetic shielding for the Xe atom at the same strategic position within an L cryptophane-A cage, under the influence of chiral potentials that represent r or l substituents outside the cage. Calculations of the Xe shielding response in the Lr and Ll diastereomeric pairs permit the prediction of the relative order of the Xe chemical shifts in solutions containing the Rl and Ll diastereomers. Where the substituent itself possesses two chiral centers, comparison of the calculated isotropic shielding responses in the Llr, Lrl, Rll, and Lrr systems, respectively, permits the prediction of the Xe spectrum of diastereomeric systems in solutions containing Llr, Rlr, Lll, and Rll systems. Assignment of the peaks observed in the experimental Xe NMR spectra is therefore possible, without having to undertake the difficult synthetic route that produces a single optically pure enantiomer.

Hyperpolarized 129Xe NMR spectroscopy can detect the presence of specific low-concentration biomolecular analytes by means of a xenon biosensor that consists of a water-soluble, targeted cryptophane-A cage that encapsulates the xenon. In this work, we use the prototypical biotinylated xenon biosensor to determine the relationship between the molecular composition of the xenon biosensor and the characteristics of protein-bound resonances. The effects of diastereomer overlap, dipole-dipole coupling, chemical-shift anisotropy, xenon exchange, and biosensor conformational exchange on the protein-bound biosensor signal were assessed. It was found that an optimal protein-bound biosensor signal can be obtained by minimizing the number of biosensor diastereomers and using a flexible linker of appropriate length. Both the line width and sensitivity of chemical shift to protein binding of the xenon biosensor were found to be inversely proportional to linker length.

Laser-enhanced (LE) 129Xe nuclear magnetic resonance (NMR) is an exceptional tool for sensing extremely small physical and chemical changes; however, the difficult mechanics of bringing polarized xenon and samples of interest together have limited applications, particularly to biological molecules. Here we present a method for accomplishing solution 129Xe biosensing based on flow (bubbling) of LE 129Xe gas through a solution in situ in the NMR probe, with pauses for data acquisition. This overcomes fundamental limitations of conventional solution-state LE 129Xe NMR, e.g., the difficulty in transferring hydrophobic xenon into aqueous environments, and the need to handle the sample to refresh LE 129Xe after an observation pulse depletes polarization. With this new method, we gained a factor of >100 in sensitivity due to improved xenon transfer to the solution and the ability to signal average by renewing the polarized xenon. Polarized xenon in biosensors was detected at very low concentrations,