Search

Post-depositional degradation of organic matter (OM) in freshwater sediments is crucial for driving the biogeochemical dynamics and influencing the consequential carbon burial. This process also often causes diagenetic alterations on paleoenvironmental proxies. Yet, mechanisms behind the degradation of sedimentary OM and the depth-related variations in stable isotope ratios can only be explained in part. Degradation of sedimentary OM in two drinking water reservoirs with contrasting trophic states (eutrophic versus mesotrophic) and catchment land-use (agriculture versus forestry) were studied with a combination of typical chemical treatments to group sedimentary organic matter into four different biochemical fractions, i.e., carbohydrates, proteins, lignins and lipids. Our results show that the degradation of sedimentary OM was mainly driven by preferential degradation of aquatic proteins and carbohydrates. In a subsequent step, lipids seemed to be degraded. This process entails more pronounced reduction of sedimentary OM and associated depth variations in δ13C and δ15N in the eutrophic reservoir of agricultural catchment, when compared to the mesotrophic reservoir with a forest dominated catchment. Moreover, changes in the relative proportions of biochemical components in sedimentary OM have more pronounced impact on δ15N isotope values relative to the ones of δ13C. Our findings also suggest that algae-derived OM is primarily responsible for the post-depositional alterations of elemental and isotopic compositions of sedimentary OM. This may be of importance for estimates of carbon sinks in lakes and reservoirs.

Parents might initially produce more offspring than they might be able to raise. However, when offspring demand exceeds their parents ' rearing capacity, parents might shift care towards the offspring which yield greater fitness returns to achieve their optimal brood size via brood reduction. Such favoritism could rely on offspring signaling traits if these inform parents about offspring quality and hence about the pay-offs of their investment. Here we investigated whether favoritism of blue tit (Cyanistes caeruleus) parents for an offspring signal (i.e., ultraviolet (UV) plumage coloration) varies with brood demand. To test this, we experimentally blocked the UV reflectance of yellow breast feathers in half of the nestlings of each brood, and then we sequentially performed two opposing brood size manipulations to vary nestling demand below or above parental rearing capacity. In reduced broods, nestlings begged overall less intensely and gained more body mass, supporting that parental rearing capacities sufficed to satisfy brood demand. Moreover, in reduced broods, UV-blocked nestlings (i.e., low-quality offspring) were fed and prey-tested more often. Yet, they begged more than control nestlings, suggesting that they were perhaps treated differently by other family members or which they may exploit parental preferences beyond actual need (at least in reduced nests). Parents flexibly shifted their feeding rate and favoritism in response to short-term changes in family size, as there was no parental preference for enlarged broods. Such flexible parental feeding rules may allow parents to gain the upper hand in parent-offspring conflict. However, we did not find evidence that parental favoritism facilitated brood reduction, at least in conditions where demand was temporally enhanced. Blue tit parents may produce more offspring than they are able to raise, and they might therefore preferentially invest in high quality offspring that are most likely to survive. Yet when parents are able to raise all their offspring they might even favor those with lowest survival probabilities to maximize reproductive success. Parents hence benefit from flexible feeding strategies for which they rely on offspring signals (i.e., colorful traits), which inform about offspring quality.

Protein complexes are defined by the three‐dimensional structure of participating binding partners. Knowledge about these structures can facilitate the design of peptidomimetics which have been applied for example, as inhibitors of protein–protein interactions (PPIs). Even though β‐sheets participate widely in PPIs, they have only rarely served as the basis for peptidomimetic PPI inhibitors, in particular when addressing intracellular targets. Here, we present the structure‐based design of β‐sheet mimetics targeting the intracellular protein β‐catenin, a central component of the Wnt signaling pathway. Based on a protein binding partner of β‐catenin, a macrocyclic peptide was designed and its crystal structure in complex with β‐catenin obtained. Using this structure, we designed a library of bicyclic β‐sheet mimetics employing a late‐stage diversification strategy. Several mimetics were identified that compete with transcription factor binding to β‐catenin and inhibit Wnt signaling in cells. The presented design strategy can support the development of inhibitors for other β‐sheet‐mediated PPIs., Starting from a 52 amino acid protein binding epitope, a bicyclic β‐hairpin structure was developed to bind the transcriptional coactivator β‐catenin. Our structure‐based design approach was supported by screening a focused library of bicyclic mimetics which was generated via late‐stage diversification. The most active bicyclic β‐hairpin shows cell‐penetration and inhibits Wnt signaling in a cell‐based assay.

In many species, offspring display conspicuous coloration already early in life, even though they might be very vulnerable to predation at this stage. However, most attention has been drawn to the conspicuous plumage displayed by adult individuals in a sexual context, while other signaling functions have been explored much less. Here, we investigated whether the yellow breast plumage of blue tit (Cyanistes caeruleus) nestlings shows patterns of condition dependence and hence signals individual quality, as has been described for adult birds. During three consecutive breeding seasons, we, therefore, explored the association between nestling body mass and three color components of the yellow breast plumage (i.e., UV chroma, carotenoid chroma, and total brightness), considering both within and among nest effects. Variation in carotenoid chroma was not related to body mass. However, UV chroma and total brightness varied with body mass on an among-nest level, suggesting that they might signal aspects of genetic quality or parental rearing capacity. Interestingly, we also found a within-nest effect of body mass on total brightness, suggesting that this is a good candidate for a condition-dependent signal within the family. Thus, other family members could rely on brightness to adjust their behavioral strategies, such as feeding behavior in parents. Our study thus reveals that certain color components of the yellow breast plumage might signal different aspects of offspring quality, and they might have a correlated signaling value across life-history stages., The study was financed by the Ministerio de Economía y Competitividad MINECO, Spain (project grants CGL2016-79390-P and PID2019-106032GB-I00 to J. Morales) funded by MCIN/AEI/https://doi.org/10.13039/ 50110 0011033 and by “ERDF A way of making Europe.” JG-C was supported by Grant BES-2017-079750 and JM by a Ramón y Cajal contract RYC-2014-15145 funded by MCIN/AEI/https://doi.org/10.13039/ 50110 0011033 and “ESF Investing in your future.”

Background: Anesthetic-induced preconditioning (AIP) with volatile anesthetics is a well-known experimental technique to protect tissues from ischemic injury or oxidative stress. Additionally, plasmatic extracellular vesicle (EV) populations and their cargo are known to be affected by AIP in vitro, and to provide organ protective properties via their cargo. We investigated whether AIP would affect the generation of EVs in an in vivo rat model. Methods: Twenty male Sprague Dawley rats received a repetitive treatment with either isoflurane or with sevoflurane for a duration of 4 or 8 weeks. EVs from blood plasma were characterized by nanoparticle tracking analysis, transmission electron microscopy (TEM) and Western blot. A scratch assay (H9C2 cardiomyoblast cell line) was performed to investigate the protective capabilities of the isolated EVs. Results: TEM images as well as Western blot analysis indicated that EVs were successfully isolated. The AIP changed the flotillin and CD63 expression on the EV surface, but not the EV concentration. The scratch assay did not show increased cell migration and/or proliferation after EV treatment. Conclusion: AIP in rats changed the cargo of EVs but had no effect on EV concentration or cell migration/proliferation. Future studies are needed to investigate the cargo on a miRNA level and to investigate the properties of these EVs in additional functional experiments.

The pig is the most widely used large animal model in Europe, with cardiovascular research being one of the main areas of application. Adequate refinement of interventional studies in this field, meeting the requirements of Russel and Burchs’ 3 R concept, can only be performed if blood-contacting medical devices are hemocompatible. Because most medical devices for cardiovascular interventional procedures are developed for humans they are tested mostly for compatibility with human blood. The aim of this study was therefore to determine whether there are differences in behavior of porcine and human platelets when they come into contact with glass, which was used as an exemplary thrombogenic material. For this purpose changes of platelet count, platelet volume and platelet expression of the activation markers CD61, CD62P and CD63 were measured using a modified chandler loop-system simulating the fluidic effects of the blood flow. Minipig and human platelets showed significant differences in number and volume, but not in activation after 4–8 min exposure to glass.

Converting biomass into value-added chemicals is of significant interest and we report an efficient hydrosilylation to convert levulinic acid to γ-valerolactone using cost-effective silanes such as PMHS and TMDS with B(C6F5)3 as catalyst. This metal free methodology works at room temperature reaching TONs and TOFs up to 16000 and 2000 h−1. Insights into the reaction mechanism are reported.

The aim of this study was to evaluate the Porophyllum ruderale effects on the liver in murine melanoma model B16-F10 and to determine the antioxidant potential. The plant’s aerial parts of the were collected in Campo Grande, MS, Brazil, dried, extracted with ethanol, ultrasonic bath and static maceration. The hydroalcoholic formulation at 100 and 200 mg/kg were submitted to antioxidant activity (DPPH) and hepatoprotective effect. Thirty-eight black mice with a mean weight of 30 g were distributed in 4 experimental groups: negative control, positive control (doxorubicin), P. ruderale at 100 mg/kg and P. ruderale at 200 mg/kg. All the animals received subcutaneous injection of 5x105 B16F10 cells on the 1st day of the experiment and were treated until the 14th day. Although a significant antioxidant activity was verified for the two concentrations analyzed, they were not sufficient to minimize the melanoma effects toxic in the mice liver as observed by histological changes and that leads us to conclude that the hydroalcoholic extract of aerial parts of P. ruderale, in the concentrations used, possesses antioxidant properties, good performance in inhibiting the proliferation of human melanoma cells, but it has no hepatoprotective effect in the melanoma cutaneous treatment. Keywords: Cutaneous Melanoma. Asteraceae. Hepatoprotective Effect. Phenolic Compounds. DPPH. Resumo O objetivo deste estudo foi avaliar os efeitos de Porophyllum ruderale no fígado em melanoma murino modelo B16-F10 e determinar o potencial antioxidante. As folhas da planta foram coletadas em Campo Grande, MS, Brasil, secas, extraídas com etanol, banho ultrassônico e maceração estática. A formulação hidroalcoólica de 100 e 200 mg/kg foi submetida à atividade antioxidante (DPPH) e ao efeito hepatoprotetor. Trinta e oito camundongos pretos com peso médio de 30 g foram distribuídos em 4 grupos experimentais: controle negativo, controle positivo (doxorrubicina), P. ruderale na concentração de 100 mg/kg e P. ruderale na concentração de 200 mg/kg. Todos os animais receberam injeção subcutânea de 5x105 células B16F10 no 1º dia de experimento até o 14º. Embora tenha sido verificada atividade antioxidante significativa para as duas concentrações analisadas, elas não foram suficientes para minimizar os efeitos tóxicos do melanoma no fígado dos camundongos, como observado nas alterações histológicas, o que nos leva a concluir que o extrato hidroalcoólico das folhas de P. ruderale, nas concentrações utilizadas, possui propriedades antioxidantes, bom desempenho na inibição da proliferação de células de melanoma humano, mas não apresenta efeito hepatoprotetor no tratamento do melanoma cutâneo. Palavras-chave: Melanoma Cutâneo. Asteraceae. Efeito Hepatoprotetor. Compostos Fenólicos. DPPH.

The acquisition of embryogenic competence involves aspects at a cellular level related to cell communication and signaling. For Araucaria angustifolia, transcriptome and proteome databases have indicated differential gene expression and protein abundance related to cell-to-cell communication during embryogenesis. The differences are mainly related to vesicular transport and regulation of plasmodesmata region-related proteins. This study aims to understand the behavior of cell-to-cell communication in cell cultures with contrasting embryogenic potential. We used two embryogenic cell lines of A. angustifolia (Responsive and Blocked) and carried out gene expression, coexpression network, immunocytochemical, and ultrastructural analyses. Eight proteins related to vesicular transport and cell-to-cell communication processes were identified with a significantly different abundance in both cell lines. One callose hydrolase (AaGHL17) was differentially expressed between the two cell lines. Accordingly, we used immunocytochemistry and found a higher intensity of callose in the Responsive, as opposed to the Blocked embryogenic cells. Transmission Electron Microscopy images highlighted a more intense vesicular activity in the Responsive cell line compared to the Blocked one. Different status of cell-to-cell intercommunication and vesicular transport is associated with somatic embryogenic competence. Vesicular transport and plasmodesmata regulation features are associated with Araucaria angustifolia somatic embryogenic responsiveness.

The goals of this study were to (a) ascertain human identity capabilities of DNA obtained from latent fingerprints that have been first environmentally insulted and then developed by the deposition of a columnar thin film (CTF), and (b) to determine if the CTF process and material are detrimental to single nucleotide polymorphism (SNP) analysis. Fingerprints were deposited on five different types of substrates and aged for one day, 7 days or 30 days while being environmentally insulted under one of the four conditions: 16.6 °C and 60% relative humidity (RH) (Condition A), 24.5 °C and 60% RH (Condition B), 35 °C and 67% RH (Condition C) and a cold condition (Condition D). Then CTF technique was then on 59% of these fingerprints. DNA samples from 805 fingerprints were extracted, quantified, subjected to manual library preparation using the Precision ID Identity Panel, and underwent high-throughput sequencing. The Ion S5™ platform was employed to sequence 124 SNP amplicons. SNPs were successfully sequenced from 802/805 samples. Total read depth was consistent across environmental conditions, and majority of samples had 100% profile completeness and 100% concordance. Anecdotally, libraries that were amplified with a higher cycle number had more ‘Major Allele Frequency’ flags compared to samples amplified with 23 cycle numbers, possibly due to stochastic effects. Neither the substrates nor the CTF process and materials inhibit downstream DNA analysis. DNA of low quality and quantity from the chosen samples can be sequenced using the Precision ID Identity Panel on the Ion S5™ platform which performed well, however, a different approach may be needed if spurious alleles are suspected.

INTRODUCTION: Due to the global epidemic of obesity, there is increasing interest in a distinct entity, called obesity-related nephropathy (ORN). Data on sustainable effects of weight reductions, with conservative, non-surgical treatment programs, on renal function in CKD patients are scarce. MATERIALS AND METHODS: We retrospectively investigated patients with CKD (eGFR ≤ 60 mL/min/1.73m2) from a non-surgical multimodality obesity treatment program over 12 months. RESULTS: We identified 17 obese patients with CKD (estimated glomerular filtration rate (eGFR) ≤ 60 mL/min at baseline). 76% were female, 41% had type II diabetes mellitus, and the mean age was 59.6 ± 8.4 years (mean ± SD). Mean serum creatinine and eGFR at baseline were 106.4 ± 17.6 µmol/L and 53.4 ± 5.8 mL/min, respectively. Mean weight and body mass index (BMI) were 134.9 ± 26.4 and 50.1 ± 10.5 kg/m2, respectively. All subjects lost weight, with average weight loss of -32.2 ± 15.1 kg (p < 0.001) by the end of 12 months (BMI at 12 months 38.1 ± 7.8 kg/m2 (-12.0 ± 6.0 kg/m2, p < 0.001). Average 12-month creatinine was 92.2 ± 23.3 µmol/L, representing a drop of 14.2 ± 15.6 µmol/L (p = 0.004). Average eGFR increased by 14.8 ± 18.0 mL/min to a 12-month value of 68.2 ± 19.3 mL/min (p = 0.002). There were no significant differences when comparing patients with and without diabetes mellitus. CONCLUSION: These results demonstrate the potential renal impact of a non-surgical multimodal obesity program on renal function in very obese patients with CKD. Weight loss intervention should be highly encouraged especially in obese CKD patients.

There is increasing evidence that individuals are consistent in the timing of their daily activities, and that individual variation in temporal behavior is related to the timing of reproduction. However, it remains unclear whether observed patterns relate to the timing of the onset of activity or whether an early onset of activity extends the time that is available for foraging. This may then again facilitate reproduction. Furthermore, the timing of activity onset and offset may vary across the breeding season, which may complicate studying the above-mentioned relationships. Here, we examined in a wild population of great tits (Parus major) whether an early clutch initiation date may be related to an early onset of activity and/or to longer active daylengths. We also investigated how these parameters are affected by the date of measurement. To test these hypotheses, we measured emergence and entry time from/into the nest box as proxies for activity onset and offset in females during the egg laying phase. We then determined active daylength. Both emergence time and active daylength were related to clutch initiation date. However, a more detailed analysis showed that the timing of activities with respect to sunrise and sunset varied throughout the breeding season both within and among individuals. The observed positive relationships are hence potentially statistical artifacts. After methodologically correcting for this date effect, by using data from the pre-egg laying phase, where all individuals were measured on the same days, neither of the relationships remained significant. Taking methodological pitfalls and temporal variation into account may hence be crucial for understanding the significance of chronotypes.

Introduction: MALDI TOF MS method is increasingly used in routine microbiological diagnostics to identify clinical strains. The result of the identification test is based on the measurement of the mass, charge and flight time of the protein ions. This makes it possible to monitor and supervise the method using a numerical score developed with statistical techniques. Aim: The study aimed to determine the stability and correctness of the mass spectrometry method. Materials and methods: To evaluate the characteristics of the method, microbial identification tests were performed using reference strains. All tests were performed as part of the MALDI TOF MS internal quality control system. Results: All reference strains (100%) were correctly identified to the species level, although the score values were not always within the reliability criteria of the results established by the producer. It was found that the mean values of the score were from 2.000 – 2.299 (49.2%) and 2.300 – 3.000 (50.0%). The coefficient of variation for control tests performed in the consecutive years ranged from 5.18 – 6.56%, which evidence of the high stability of the method. For individual species, reproducibility precision over the 8 years ranged from 2.89% (n = 13) for Enterococcus faecalis ATCC 51299 to 7.02% (n = 28) for Klebsiella pneumoniae ATCC 700603, which proves the high precision of measurements. Conclusions: The mass spectrometry method is characterized by very high stability and correctness. The intra-laboratory quality control system using reference strains is a useful tool for monitoring and supervising the method and laboratory personnel competency performing identification tests during routine microbiological work.

A novel, versatile approach for the synthesis of unsymmetrical 3,3'-diindolylmethanes (DIMs) with a quaternary carbon center has been developed via iodine-catalyzed coupling of trifluoromethyl(indolyl)phenylmethanols with indoles. In contrast to previously reported methods, the new procedure is characterized by chemoselectivity, mild conditions, high yields, and scalability to obtain gram amounts for biological studies. Selected compounds were found to display affinity for cannabinoid receptors, which are promising drug targets for the treatment of inflammatory and neurodegenerative diseases.

Vascular smooth muscle (VSM) cell phenotypic expression and autophagic state are dynamic responses to stress. Vascular pathologies, such as hypoxemia and ischemic injury, induce a synthetic VSM phenotype and autophagic flux resulting in a loss of vascular integrity and VSM cell death respectfully. Both clinical pilot and experimental stroke studies demonstrate that sphingosine-1-phosphate receptor (S1PR) modulation improves stroke outcome; however, specific mechanisms associated with a beneficial outcome at the level of the cerebrovasculature have not been clearly elucidated. We hypothesized that ozanimod, a selective S1PR type 1 ligand, will attenuate VSM synthetic phenotypic expression and autophagic flux in primary human brain VSM cells following acute hypoxia plus glucose deprivation (HGD; in vitro ischemic-like injury) exposure. Cells were treated with ozanimod and exposed to normoxia or HGD. Crystal violet staining, standard immunoblotting, and immunocytochemical labeling techniques assessed cellular morphology, vacuolization, phenotype, and autophagic state. We observed that HGD temporally decreased VSM cell viability and concomitantly increased vacuolization, both of which ozanimod reversed. HGD induced a simultaneous elevation and reduction in levels of pro- and antiautophagic proteins respectfully, and ozanimod attenuated this response. Protein levels of VSM phenotypic biomarkers, smoothelin and SM22, were decreased following HGD. Furthermore, we observed an HGD-induced epithelioid and synthetic morphological appearance accompanied by disorganized cytoskeletal filaments, which was rescued by ozanimod. Thus, we conclude that ozanimod, a selective S1PR1 ligand, protects against acute HGD-induced phenotypic switching and promotes cell survival, in part, by attenuating HGD-induced autophagic flux thus improving vascular patency in response to acute ischemia-like injury.

Merkel cell carcinoma (MCC) is a rare, but aggressive skin cancer the incidence of which has increased significantly in recent years. The majority of MCCs have incorporated Merkel cell polyomavirus (VP-MCC) while the remainder are virus-negative (VN-MCC). Although a variety of therapeutic options have shown promise in treating MCC, there remains a need for additional therapeutics as well as probes for better understanding MCC. A high-throughput screening campaign was used to assess the ability of > 25,000 synthetic and natural product compounds as well as > 20,000 natural product extracts to affect growth and survival of VN-MCC and VP-MCC cell lines. Sixteen active compounds were identified that have mechanisms of action reported in the literature along with a number of compounds with unknown mechanisms. Screening results with pure compounds suggest a range of potential targets for MCC including DNA damage, inhibition of DNA or protein synthesis, reactive oxygen species, and proteasome inhibition as well as NFκB inhibition while also suggesting the importance of zinc and/or copper binding. Many of the active compounds, particularly some of the natural products, have multiple reported targets suggesting that this strategy might be a particularly fruitful approach. Processing of several active natural product extracts resulted in the identification of additional MCC-active compounds. Based on these results, further investigations focused on natural products sources, particularly of fungal origin, are expected to yield further potentially useful modulators of MCC.

Anthropogenic stressors, such as artificial light at night (ALAN), increasingly affect the sleep behaviour and physiology of wild birds, particularly in areas where human activity is prevalent. To understand the consequences of the resulting sleep deprivation, it is essential to investigate whether the effects of sleep deprivation on cognitive performance, ob-served in humans, also occur in birds. Here, we studied the impact of sleep deprivation, induced by intermittent expo-sure to ALAN, on inhibitory control, vigilance behaviour, and exploratory behaviour in great tits. Furthermore, we hypothesised that the effect of ALAN could depend on an individual's natural sleep duration and the timing of sleep. To achieve these goals, we measured emergence and entry times from/into the nest box in the wild, before capturing the great tits. In captivity, half of the birds were exposed to intermittent ALAN, and cognitive performance was assessed the following morning for all birds. ALAN-exposed birds were less successful on the detour reach task and when they started pecking at the test tube, they pecked more often. However, neither of the effects was related to the natural sleep duration or timing, in contrast to our hypothesis, and there were no differences between the ALAN-exposed and non-exposed group in vigilance and exploratory behaviour. Thus, even one night of exposure to ALAN can negatively affect cognitive performance in wild birds, possibly with negative effects on their performance and survival.

As environmental conditions fluctuate across years, seasonal migrants must determine where and when to move without comprehensive knowledge of conditions beyond their current location. Animals can address this challenge by following cues in their local environment to vary behaviour in response to current conditions, or by moving based on learned or inherited experience of past conditions resulting in fixed behaviour across years.It is often claimed that long‐distance migrants are more fixed in their migratory behaviour because as distance between breeding and wintering areas increases, reliability of cues to predict distant and future conditions decreases. While supported by some population‐level studies, the influence of migration distance on behavioural variation is seldom examined on an individual level.Lesser black‐backed gulls Larus fuscus are generalist seabirds that use a diversity of migration strategies. Using high‐resolution multi‐year GPS tracking data from 82 individuals from eight colonies in Western Europe, we quantified inter‐ and intra‐individual variation in non‐breeding distributions, winter site fidelity, migration routes and timing of migration, with the objectives of determining how much variation lesser black‐backed gulls have in their migratory behaviour and examining whether variation changes with migration distance.We found that intra‐individual variation was significantly lower than variation between individuals for non‐breeding distributions, winter site fidelity, migration routes and timing of migration, resulting in consistent individual strategies for all behaviours examined. Yet, intra‐individual variation ranged widely among individuals (e.g. winter site overlap: 0–0.91 out of 1; migration timing: 0–192 days), and importantly, individual differences in variation were not related to migration distance.The apparent preference for maintaining a consistent strategy, present in even the shortest distance migrants, suggests that familiarity may be more advantageous than exactly tracking current environmental conditions. Yet, variation in behaviour across years was observed in many individuals and could be substantial. This suggests that individuals, irrespective of migration distance, have the capacity to adjust to current conditions within the broad confines of their individual strategies, and occasionally, even change their strategy., This study uses multi‐year high resolution GPS tracks of lesser black‐backed gulls to quantify variation in migration behaviour at the individual‐level and along broad range migration distances to demonstrate that long‐distance migrants are no more fixed in their migratory behaviours than short distance migrants, contrasting with findings from population‐level studies.

Araucaria angustifolia is classified as a critically endangered species by the International Union for Conservation of Nature. This threat is worsened by the inefficiency of methods for ex-situ conservation and propagation. In conifers, somatic embryogenesis (SE) associated with cryopreservation is an efficient method to achieve germplasm conservation and mass clonal propagation. However, the efficiency of SE is highly dependent on genotype responsivity to the artificial stimulus used in vitro during cell line proliferation and later during somatic embryo development. In this study, we evaluated the activity of antioxidant enzymes and characterized mitochondrial functions during the proliferation of embryogenic cells of A. angustifolia responsive (SE1) and non-responsive (SE6) to the development of somatic embryos. The activities of the antioxidant enzymes GR (EC 1.6.4.2), MDHAR (EC 1.6.5.4), and POX (EC 1.11.1.7) were increased in SE1 culture, while in SE6 culture, only the activity of DHAR (EC 1.8.5.1) was significantly higher. Additionally, SE6 culture presented a higher number of mitochondria, which agreed with the increased rate of oxygen consumption compared to responsive SE1 culture; however, the mitochondrial volume was lower. Although the ATP levels did not differ, the NAD(P)H levels were higher in SE1 cells. NDs, AOX, and UCP were less active in responsive SE1 than in non-responsive cells. Our results show significant differences between SE1 and SE6 embryogenic cells regarding mitochondrial functions and antioxidant enzyme activities, which may be intrinsic to the in vitro proliferation phase of both cell lines, possessing a crucial role for the induction of in vitro maturation process.

International audience; Background: Ofatumumab, a subcutaneous anti-CD20 monoclonal antibody, selectively depletes B cells. Teriflunomide, an oral inhibitor of pyrimidine synthesis, reduces T-cell and B-cell activation. The relative effects of these two drugs in patients with multiple sclerosis are not known.Methods: In two double-blind, double-dummy, phase 3 trials, we randomly assigned patients with relapsing multiple sclerosis to receive subcutaneous ofatumumab (20 mg every 4 weeks after 20-mg loading doses at days 1, 7, and 14) or oral teriflunomide (14 mg daily) for up to 30 months. The primary end point was the annualized relapse rate. Secondary end points included disability worsening confirmed at 3 months or 6 months, disability improvement confirmed at 6 months, the number of gadolinium-enhancing lesions per T1-weighted magnetic resonance imaging (MRI) scan, the annualized rate of new or enlarging lesions on T2-weighted MRI, serum neurofilament light chain levels at month 3, and change in brain volume.Results: Overall, 946 patients were assigned to receive ofatumumab and 936 to receive teriflunomide; the median follow-up was 1.6 years. The annualized relapse rates in the ofatumumab and teriflunomide groups were 0.11 and 0.22, respectively, in trial 1 (difference, -0.11; 95% confidence interval [CI], -0.16 to -0.06; P

Protein complex formation depends on the interplay between preorganization and flexibility of the binding epitopes involved. The design of epitope mimetics typically focuses on stabilizing a particular bioactive conformation, often without considering conformational dynamics, which limits the potential of peptidomimetics against challenging targets such as transcription factors. We developed a peptide‐derived inhibitor of the NF‐Y transcription factor by first constraining the conformation of an epitope through hydrocarbon stapling and then fine‐tuning its flexibility. In the initial set of constrained peptides, a single non‐interacting α‐methyl group was observed to have a detrimental effect on complex stability. Biophysical characterization revealed how this methyl group affects the conformation of the peptide in its bound state. Adaption of the methylation pattern resulted in a peptide that inhibits transcription factor assembly and subsequent recruitment to the target DNA., Set Me free: A peptide‐derived inhibitor of the NF‐Y transcription factor was developed by constraining the conformation of an epitope through hydrocarbon stapling and then fine‐tuning its flexibility. In the initial set of constrained peptides, a non‐interacting α‐methyl group was observed to have a detrimental effect on complex stability. Adaption of the methylation pattern gave a peptide that inhibits transcription factor assembly and subsequent DNA binding.

Uranium–uranium ( 234 U– 238 U) disequilibrium dating can determine the age of secondary carbonates over greater time intervals than the well-established 230 Th– 234 U dating method. Yet it is rarely applied due to unknowns in the initial δ234 U ( δ234 U i ) value, which result in significant age uncertainties. In order to understand the δ234 U i in Devils Hole 2 cave, Nevada, we have determined 110 δ234 U i values from phreatic calcite using 230 Th– 234 U disequilibrium dating. The sampled calcite was deposited in Devils Hole 2 between 4 and 590 ka, providing a long-term look at δ234 U i variability over time. We then performed multi-linear regression among the δ234 U i values and correlative δ18 O and δ13 C values. The regression can be used to estimate the δ234 U i value of Devils Hole calcite based upon its measured δ18 O and δ13 C values. Using this approach and the measured present-day δ234 U values of Devils Hole 2 calcite, we calculated 110 independent 234 U– 238 U ages. In addition, we used newly measured δ18 O, δ13 C, and present-day δ234 U values to calculate 10 234 U– 238 U ages that range between 676 and 731 ka, thus allowing us to extend the Devils Hole chronology beyond the 230 Th– 234 U-dated chronology while maintaining an age precision of ∼ 2 %. Our results indicate that calcite deposition at Devils Hole 2 cave began no later than 736 ± 11 kyr ago. The novel method presented here may be applied to future speleothem studies in similar hydrogeological settings, given appropriate calibration studies.

Biomass and waste derived fuels contain large amounts of sodium, potassium, and chlorine that form NaCl and KCl, that is, compounds that cause operational problems, such as slagging, fouling, and high-temperature corrosion. Therefore, alkali chlorides are the main reasons that explain why steam parameters are less advanced and efficient in biomass and waste-based power generation when compared to coal. These problems can be mitigated by introducing sulphur into the system to form alkali sulphates that are not as problematic on steel surfaces as alkali chlorides. However, the alkali sulphation process in realistic combustion environments needs further exploration. Thus, new diagnostic methods for in-situ monitoring of alkali sulphation kinetics in combustion systems are required. In this work, the simultaneous monitoring of KCl and KOH concentrations in a pilot-scale combustor using Collinear Photofragmentation and Atomic Absorption Spectroscopy (CPFAAS) during stationary and transient operation of the combustor, is introduced. The CPFAAS information is complemented by monitoring SO2 and HCl concentrations using Fourier-transform infrared spectroscopy (FTIR). The temporal performance of the system is demonstrated by measuring the temporal combustor response curves for KCl sulphation for different Cl/K ratios during rapid changes in gaseous SO2 concentrations. The temporal concentration curves obtained imply that the Cl/K ratio has a significant impact on the temporal alkali sulphation behaviour. The measurement system described enables further exploration of K–Cl–S chemistry in realistic large-scale power plant environments.

Trastuzumab-emtansine (T-DM1) is an antibody-drug conjugate (ADC) that efficiently delivers a highly potent microtubule inhibitor to HER2 overexpressing cells. Herein, we utilize HER2 transformed human mammary epithelial cells (HME2) to demonstrate in vitro and in vivo response and recurrence upon T-DM1 treatment. Continuous in vitro dosing of HME2 cells with T-DM1 failed to produce a spontaneously resistant cell line. However, induction of epithelial–mesenchymal transition (EMT) via pretreatment with TGF-β1 was capable of promoting emergence of T-DM1-resistant (TDM1R) cells. Flow cytometric analyses indicated that induction of EMT decreased trastuzumab binding, prior to overt loss of HER2 expression in TDM1R cells. Kinome analyses of TDM1R cells indicated increased phosphorylation of ErbB1, ErbB4, and FGFR1. TDM1R cells failed to respond to the ErbB kinase inhibitors lapatinib and afatinib, but they acquired sensitivity to FIIN4, a covalent FGFR kinase inhibitor. In vivo, minimal residual disease (MRD) remained detectable via bioluminescent imaging following T-DM1-induced tumor regression. Upon cessation of the ADC, relapse occurred and secondary tumors were resistant to additional rounds of T-DM1. These recurrent tumors could be inhibited by FIIN4. Moreover, ectopic overexpression of FGFR1 was sufficient to enhance tumor growth, diminish trastuzumab binding, and promote recurrence following T-DM1-induced MRD. Finally, patient-derived xenografts from a HER2+ breast cancer patient who had progressed on trastuzumab failed to respond to T-DM1, but tumor growth was significantly inhibited by FIIN4. Overall, our studies strongly support therapeutic combination of TDM1 with FGFR-targeted agents in HER2+ breast cancer.

The focus of this paper is on effects of chlorine and sulfur on coal ash deposition rates, under practically relevant but systematically controlled combustion conditions. This problem is important, not so much for coal, but to understand and predict deposition rates for biomass combustion where chlorine contents can be high. To this end, ash deposition rates on a controlled temperature surface were measured for controlled amounts of chlorine and sulfur added to a pulverized coal, doped with potassium and burned in a 100 kW rated combustion rig. Previous work with 35 tests on 11 coal, biomass and petroleum coke fuels burned under a range of operating conditions had strongly suggested that the deposition rate of the tightly bound inside deposits was independent of the ash aerosol composition, and depended only on PM1 in the flue gas. The loosely bound outside deposition rate was dependent primarily on the total alkali content in the flue gas. The new results using chlorine added to the fuel (in the form of ammonium chloride) required these previous conclusions to be drastically revised. They showed that chlorine, not alkali alone, had large effects on the deposition rate of the inside deposits, which now were orders of magnitude higher than without chlorine addition, and did not fit previous (multi-fuel) correlations with PM1. Sulfur addition, together with chlorine, did not affect deposition rates much, although it did lower the chlorine content of the deposit. These results are interpreted in terms of the ash aerosol size segregated composition, which was also measured, and potential sulfation reactions within the deposit.

Purpose: The aim of this study was to describe the surgical repair of a rhegmatogenous retinal detachment (RRD) with a posterior break in a child with retinoblastoma (RB). Methods: Retrospective case report and review of the English language literature. Retrospective review of an 11-month-old male with bilateral retinoblastoma who developed a RRD with a posterior retinal break in his better-seeing eye after treatment with cryotherapy. A review of all published cases to date of RRD in patients with RB is presented. Results: The patient underwent a posterior segmental scleral buckle without subretinal fluid drainage with successful reattachment of the retina and no extraocular extension of RB. Conclusions: RRDs in RB patients may be successfully repaired with anatomic success and no extraocular tumor extension. Even for patients with a posterior break, a segmental scleral buckle without drainage of subretinal fluid is a viable option and long-term excellent vision is a possible outcome.

Background: Small studies have recently suggested that there are specific plasma metabolic signatures in chronic obstructive pulmonary disease (COPD), but there have been no large comprehensive study of metabolomic signatures in COPD that also integrate genetic variants. Materials and Methods: Fresh frozen plasma from 957 non-Hispanic white subjects in COPDGene was used to quantify 995 metabolites with Metabolon's global metabolomics platform. Metabolite associations with five COPD phenotypes (chronic bronchitis, exacerbation frequency, percent emphysema, post-bronchodilator forced expiratory volume at one second [FEV1]/forced vital capacity [FVC], and FEV1 percent predicted) were assessed. A metabolome-wide association study was performed to find genetic associations with metabolite levels. Significantly associated single-nucleotide polymorphisms were tested for replication with independent metabolomic platforms and independent cohorts. COPD phenotype-driven modules were identified in network analysis integrated with genetic associations to assess gene-metabolite-phenotype interactions. Results: Of metabolites tested, 147 (14.8%) were significantly associated with at least 1 COPD phenotype. Associations with airflow obstruction were enriched for diacylglycerols and branched chain amino acids. Genetic associations were observed with 109 (11%) metabolites, 72 (66%) of which replicated in an independent cohort. For 20 metabolites, more than 20% of variance was explained by genetics. A sparse network of COPD phenotype-driven modules was identified, often containing metabolites missed in previous testing. Of the 26 COPD phenotype-driven modules, 6 contained metabolites with significant met-QTLs, although little module variance was explained by genetics. Conclusion: A dysregulation of systemic metabolism was predominantly found in COPD phenotypes characterized by airflow obstruction, where we identified robust heritable effects on individual metabolite abundances. However, network analysis, which increased the statistical power to detect associations missed previously in classic regression analyses, revealed that the genetic influence on COPD phenotype-driven metabolomic modules was modest when compared with clinical and environmental factors.

New PCNPy pincer nickel complexes have been synthesized through a short synthetic route. Incorporating pyridine as the nitrogen side arm facilitated the C–H activation in the PCN ligand and allowed the cyclometallation with nickel to take place at room temperature. Pyridine also enhanced the stability of β-hydrogen-containing alkyl complexes. Also, the symmetric NCN nickel complex with pyridine side arms was successfully obtained giving a rare example of such type of complexes to be prepared through direct C–H activation. Furthermore, preliminary results showed that the (PCNPy)Ni–Br is active in Kumada coupling reactions particularly the coupling of aryl halides with aryl Grignard reagents.

This report describes local administration of submicron particle paclitaxel (SPP) (NanoPac®: ~ 800-nm-sized particles with high relative surface area with each particle containing ~ 2 billion molecules of paclitaxel) in preclinical models and clinical trials evaluating treatment of carcinomas. Paclitaxel is active in the treatment of epithelial solid tumors including ovarian, peritoneal, pancreatic, breast, esophageal, prostate, and non-small cell lung cancer. SPP has been delivered directly to solid tumors, where the particles are retained and continuously release the drug, exposing primary tumors to high, therapeutic levels of paclitaxel for several weeks. As a result, tumor cell death shifts from primarily apoptosis to both apoptosis and necroptosis. Direct local tumoricidal effects of paclitaxel, as well as stimulation of innate and adaptive immune responses, contribute to antineoplastic effects. Local administration of SPP may facilitate tumor response to systemically administered chemotherapy, targeted therapy, or immunotherapy without contributing to systemic toxicity. Results of preclinical and clinical investigations described here suggest that local administration of SPP achieves clinical benefit with negligible toxicity and may complement standard treatments for metastatic disease. Graphical abstract

Elevated dissolved iron concentrations in the methanic zone are typical geochemical signatures of rapidly accumulating marine sediments. These sediments are often characterized by co-burial of iron oxides with recalcitrant aromatic organic matter of terrigenous origin. Thus far, iron oxides are predicted to either impede organic matter degradation, aiding its preservation, or identified to enhance organic carbon oxidation via direct electron transfer. Here, we investigated the effect of various iron oxide phases with differing crystallinity (magnetite, hematite, and lepidocrocite) during microbial degradation of the aromatic model compound benzoate in methanic sediments. In slurry incubations with magnetite or hematite, concurrent iron reduction, and methanogenesis were stimulated during accelerated benzoate degradation with methanogenesis as the dominant electron sink. In contrast, with lepidocrocite, benzoate degradation, and methanogenesis were inhibited. These observations were reproducible in sediment-free enrichments, even after five successive transfers. Genes involved in the complete degradation of benzoate were identified in multiple metagenome assembled genomes. Four previously unknown benzoate degraders of the genera Thermincola (Peptococcaceae, Firmicutes), Dethiobacter (Syntrophomonadaceae, Firmicutes), Deltaproteobacteria bacteria SG8_13 (Desulfosarcinaceae, Deltaproteobacteria), and Melioribacter (Melioribacteraceae, Chlorobi) were identified from the marine sediment-derived enrichments. Scanning electron microscopy (SEM) and catalyzed reporter deposition fluorescence in situ hybridization (CARD-FISH) images showed the ability of microorganisms to colonize and concurrently reduce magnetite likely stimulated by the observed methanogenic benzoate degradation. These findings explain the possible contribution of organoclastic reduction of iron oxides to the elevated dissolved Fe2+ pool typically observed in methanic zones of rapidly accumulating coastal and continental margin sediments.

Coronaviruses (CoVs) infect both humans and animals. In humans, CoVs can cause respiratory, kidney, heart, brain, and intestinal infections that can range from mild to lethal. Since the start of the 21st century, three β‐coronaviruses have crossed the species barrier to infect humans: severe‐acute respiratory syndrome (SARS)‐CoV‐1, Middle East respiratory syndrome (MERS)‐CoV, and SARS‐CoV‐2 (2019‐nCoV). These viruses are dangerous and can easily be transmitted from human to human. Therefore, the development of anticoronaviral therapies is urgently needed. However, to date, no approved vaccines or drugs against CoV infections are available. In this review, we focus on the medicinal chemistry efforts toward the development of antiviral agents against SARS‐CoV‐1, MERS‐CoV, SARS‐CoV‐2, targeting biochemical events important for viral replication and its life cycle. These targets include the spike glycoprotein and its host‐receptors for viral entry, proteases that are essential for cleaving polyproteins to produce functional proteins, and RNA‐dependent RNA polymerase for viral RNA replication.

Extraction of olive oil gives rise to large quantities of pomace and liquid effluents, since on average only 21% of the weight of the olive corresponds to oil, the remaining 79% consists of water, bark, pulp and stone. With the intention to make available new forms of use of this residue, this research was proposed, with aimed to optimize the extraction of phenolic compounds from olive pomace resulting from oil extraction using methanolic extracts. The analysis of phenolic compounds (TPC) and the evaluation of the antioxidant activity (AA) were performed by spectrophotometry, and the individual phenols were carried out by LC-ESI-qTOF-MS. The data were evaluated by the application of the response surface methodology (RSM). The condition that promoted the highest TPC in an extract was using 40% methanol, 70 °C and 180 minutes (extract 7). The highest AA was in the extract obtained with 40% methanol, 45 °C and 180 minutes (extract 5). The highest individual phenol sum (IPS) was in the extract with 80% methanol, 45 oC and 180 minutes (extract 6). Therefore, it is possible to conclude that the RSM was an interesting tool to measure the best conditions for extraction of phenolic compounds from olive pomace.