Discovery of potent small-molecule inhibitors of lipoprotein(a) formation.

Lipoprotein(a) (Lp(a)), an independent, causal cardiovascular risk factor, is a lipoprotein particle that is formed by the interaction of a low-density lipoprotein (LDL) particle and apolipoprotein(a) (apo(a))1,2. Apo(a) first binds to lysine residues of apolipoprotein B-100 (apoB-100) on LDL through the Kringle IV (KIV) 7 and 8 domains, before a disulfide bond forms between apo(a) and apoB-100 to create Lp(a) (refs. 3-7). Here we show that the first step of Lp(a) formation can be inhibited through small-molecule interactions with apo(a) KIV7-8. We identify compounds that bind to apo(a) KIV7-8, and, through chemical optimization and further application of multivalency, we create compounds with subnanomolar potency that inhibit the formation of Lp(a). Oral doses of prototype compounds and a potent, multivalent disruptor, LY3473329 (muvalaplin), reduced the levels of Lp(a) in transgenic mice and in cynomolgus monkeys. Although multivalent molecules bind to the Kringle domains of rat plasminogen and reduce plasmin activity, species-selective differences in plasminogen sequences suggest that inhibitor molecules will reduce the levels of Lp(a), but not those of plasminogen, in humans. These data support the clinical development of LY3473329-which is already in phase 2 studies-as a potent and specific orally administered agent for reducing the levels of Lp(a).

Determinants of Frame Running Capacity in Athletes With Cerebral Palsy to Improve Training Routines and Classification Strategies: A Cross-sectional Observational Study.

OBJECTIVES: The aim of the study were to (1) investigate what physical and physiological parameters are most important for Frame Running capacity, a parasport for individuals with ambulatory difficulties, and (2) determine whether Frame Running capacity can be predicted in athletes with cerebral palsy. DESIGN: Athletes with cerebral palsy ( N = 62, Gross Motor Classification System I-V; 2/26/11/21/2) completed a 6-min Frame Running test. Before the 6-min Frame Running test, muscle thickness, passive range of motion (hip, knee, ankle), selective motor control, and spasticity (hip, knee, ankle) were measured in both legs. In total, 54 variables per individual were included. Data were analyzed using correlations, principal component analysis, orthogonal partial least square regression, and variable importance in projection analysis. RESULTS: The mean 6-min Frame Running test distance was 789 ± 335 m and decreased with motor function severity. The orthogonal partial least square analysis revealed a modest degree of covariance in the variables analyzed and that the variance in the 6-min Frame Running test distance could be predicted with 75% accuracy based on all the variables measured. Variable importance in projection analysis indicated hip and knee extensor spasticity (negative effect), and muscle thickness (positive effect) arose as the most important factors contributing to Frame Running capacity. CONCLUSIONS: These results are an important resource to enable optimization of training regimes to improve Frame Running capacity and contribute to evidence-based and fair classification for this parasport.

Determining occupational accidents baseline ratios by considering a synthetic population: The case of Spain.

In most countries, a government agency or collaborating organization gathers information on occupational accidents. Comparisons based on a single factor such as autonomous community, activity sector or others, often leads to contradictory conclusions. The use of this information for comparison is not immediate because the different characteristics considered give place to different possible comparisons. The elaboration of a single baseline for each set of characteristics is addressed. The method proposed comes from the data available in Spain but could be applied to other cases. The method consists of: (1) selecting factors-those selected are age, sex, autonomous community and activity; (2) the generation of a synthetic population based on data from a survey and general proportions by applying the Optimal Representative Sample Weighting (rsw); and (3) the prediction of the accidents ratio for each set of characteristic by using a XGBoost decision trees ensemble. The results confirm the appropriateness of the method.

Enterobacteria impair host p53 tumor suppressor activity through mRNA destabilization.

Increasing evidence highlights the role of bacteria in the physiopathology of cancer. However, the underlying molecular mechanisms remains poorly understood. Several cancer-associated bacteria have been shown to produce toxins which interfere with the host defense against tumorigenesis. Here, we show that lipopolysaccharides from Klebsiella pneumoniae and other Enterobacteria strongly inhibit the host tumor suppressor p53 pathway through a novel mechanism of p53 regulation. We found that lipopolysaccharides destabilize TP53 mRNA through a TLR4-NF-κB-mediated inhibition of the RNA-binding factor Wig-1. Importantly, we show that K. pneumoniae disables two major tumor barriers, oncogene-induced DNA damage signaling and senescence, by impairing p53 transcriptional activity upon DNA damage and oncogenic stress. Furthermore, we found an inverse correlation between the levels of TLR4 and p53 mutation in colorectal tumors. Hence, our data suggest that the repression of p53 by Enterobacteria via TLR4 alleviates the selection pressure for p53 oncogenic mutations and shapes the genomic evolution of cancer.

Effects of the COVID-19 Health Crisis on Sports Practice, Life Quality, and Emotional Status in Spanish High-Performance Athletes.

Spain is one of the many countries highly affected by the COVID-19 crisis, establishing very restrictive measures with a complete lockdown for more than 3 months. This situation forced the complete closure of sport practice and national or international competitions, leading to a negative impact on physical and psychological health of high-performance athletes. Therefore, the objectives of this study were (a) to determine the effects of the COVID-19 health crisis on Spanish high-performance athletes in terms of sports practice, life quality, and emotional state and (b) to identify the profile with the greatest difficulties during and after the lockdown. A sample of 130 high-performance athletes aged between 18 and 34 years (67 women and 63 men) participated in this study (83.1% achieved a medal in National-International elite competitions; 86.9% were considered student-athletes). Measures included socio-demographic data through a 5-dimension ad hoc survey: physical activity and exercise using an adapted version from the International Physical Activity Questionnaire (IPAQ); health status and limitations using an adapted version of SF-12 Health Questionnaire; Perceived stress (Short-PSS); and Mood States (29-item POMS). All participants have shown a significant decrease pre-post-lockdown in both health and performance perception, especially in women, individual athletes, medalists, and student-athletes. Strong limitations of training, attention, and motivation as well as a moderate negative emotional state during lockdown were reported, in women, individual athletes, medalists, and student-athletes. Even with an improved emotional state and energy level in the post-lockdown period, moderate-to-high stress scores were reported by women and medalists. Our findings highlight the importance of paying attention to the physical and psychological health of elite athletes on three profiles: team athletes (due to social distance), student-athletes (dual-career issues), and women athletes (prevalence of implicit gender inequalities in sport).

Effects of Whole-Body Vibration Training on Body Composition, Cardiometabolic Risk, and Strength in the Population Who Are Overweight and Obese: A Systematic Review With Meta-analysis.

OBJECTIVES: To assess the effects of whole-body vibration training (WBVT) on body composition, metabolic and cardiovascular risk variables, and lower limb strength in participants who are overweight/obese. DATA SOURCES: A systematic review with meta-analysis was conducted in 3 databases (PubMed-MEDLINE, Web of Science, and Cochrane Library) from inception through to January 26, 2020. STUDY SELECTION: Studies analyzing the effect of WBVT on body composition variables, metabolic profile, blood pressure, heart rate, and lower limb strength in the population who are overweight/obese, with interventions of a minimum length of 2 weeks were included. DATA EXTRACTION: After applying the inclusion and exclusion criteria, 23 studies involving 884 participants who were obese/overweight (experimental group: 543; weight=79.9 kg; body mass index (BMI) =31.3 kg/m2, obesity class I according to World Health Organization) were used in the quantitative analysis. The sex of the participants involved in the studies were as follows: (1) 17 studies included only female participants; (2) 1 study included only boys, and (3) 5 studies included both sexes. Meta-analysis, subgroup analysis, and meta-regression methods were used to calculate the mean difference and standardized mean difference (SMD; ± 95% confidence intervals [CIs]) as well as to analyze the effects of pre-post intervention WBVT and differences from control groups. DATA SYNTHESIS: WBVT led to a significant decrease in fat mass (-1.07 kg, not clinically significant). In addition, WBVT reduced systolic blood pressure (-7.01 mmHg, clinically significant), diastolic blood pressure (-1.83 mmHg), and heart rate (-2.23 bpm), as well as increased the lower extremity strength (SMD=0.63; range, 0.40-0.86). On the other hand, WBVT did not modify the weight, BMI, muscle mass, cholesterol, triglycerides, or glucose. CONCLUSIONS: WBVT could be an effective training modality to reduce blood pressure (clinically relevant) and resting heart rate. In addition, WBVT led to improved lower limb strength. However, these findings were not consistent with significant improvements on other variables associated with metabolic syndrome (body composition, cholesterol, triglycerides, glucose).

Variability in vastus lateralis fiber type distribution, fiber size, and myonuclear content along and between the legs.

Human skeletal muscle characteristics such as fiber type composition, fiber size, and myonuclear content are widely studied in clinical and sports-related contexts. Being aware of the methodological and biological variability of the characteristics is a critical aspect in study design and outcome interpretation, but comprehensive data on the variability of morphological features in human skeletal muscle are currently limited. Accordingly, in the present study, m. vastus lateralis biopsies (10 per subject) from young and healthy individuals, collected in a systematic manner, were analyzed for various characteristics using immunohistochemistry (n = 7) and SDS-PAGE (n = 25). None of the analyzed parameters, fiber type % (FT%), type I and II fiber cross-sectional area (fCSA), percentage fiber type area (fCSA%), myosin heavy chain composition (MyHC%), type IIX content, myonuclear content, or myonuclear domain, varied in a systematic manner longitudinally along the muscle or between the two legs. The average within-subject coefficient of variation for FT%, fCSA, fCSA%, and MyHC% ranged between 13% and 18% but was only 5% for fiber-specific myonuclear content, which reduced the variability for myonuclear domain size to 11%-12%. Pure type IIX fibers and type IIX MyHC were randomly distributed and present in <24% of the analyzed samples, with the average content being 0.1% and 1.1%, respectively. In conclusion, leg or longitudinal orientation does not seem to be an important aspect to consider when investigating human vastus lateralis characteristics. However, single muscle biopsies should preferably not be used when studying fiber type- and fiber size-related aspects, given the notable sample-to-sample variability.NEW & NOTEWORTHY This study provides a comprehensive analysis of the variability of key human skeletal muscle fiber characteristics in multiple sites along and between the m. vastus lateralis of healthy and active individuals. We found a notable but nonsystematic variability in fiber type and size, whereas myonuclear content was distinctively less variable, and the prevalence of type IIX fibers was random and very low. These data are important to consider when designing and interpreting studies including m. vastus lateralis biopsies.

Early accentuated muscle hypertrophy is strongly associated with myonuclear accretion.

The current study explored whether the marked hypertrophic response noted with a short-term unilateral concurrent exercise paradigm was associated with more prominent changes in myonuclei accretion, ribosome biogenesis, and capillarization compared with resistance exercise alone (RE). Ten men (age 25 ± 4 yr) performed aerobic and resistance exercise (AE + RE) for one leg while the other leg did RE. Muscle biopsies were obtained before and after 5 wk of training and subjected to fiber-type specific immunohistochemical analysis, and quantification of total RNA content and mRNA/rRNA transcript abundance. Type II fiber cross-sectional area (CSA) increased with both AE + RE (22%) and RE (16%), while type I fiber CSA increased mainly with AE + RE (16%). The change score tended to differ between legs for type I CSA (P = 0.099), and the increase in smallest fiber diameter was greater in AE + RE than RE (P = 0.029). The number of nuclei per fiber increased after AE + RE in both fiber types, and this increase was greater (P = 0.027) than after RE. A strong correlation was observed between changes in number of nuclei per fiber and fiber CSA in both fiber types, for both AE + RE and RE (r > 0.8, P < 0.004). RNA content increased after AE + RE (24%, P = 0.019), but the change-scores did not differ across legs. The capillary variables generally increased in both fiber types, with no difference across legs. In conclusion, the accentuated hypertrophic response to AE + RE was associated with more pronounced myonuclear accretion, which was strongly correlated with the degree of fiber hypertrophy. This suggests that myonuclear accretion could play a role in facilitating muscle hypertrophy also during very short training periods.

Thermal Proteome Profiling Identifies Oxidative-Dependent Inhibition of the Transcription of Major Oncogenes as a New Therapeutic Mechanism for Select Anticancer Compounds.

Identification of the molecular mechanism of action (MoA) of bioactive compounds is a crucial step for drug development but remains a challenging task despite recent advances in technology. In this study, we applied multidimensional proteomics, sensitivity correlation analysis, and transcriptomics to identify a common MoA for the anticancer compounds RITA, aminoflavone (AF), and oncrasin-1 (Onc-1). Global thermal proteome profiling revealed that the three compounds target mRNA processing and transcription, thereby attacking a cancer vulnerability, transcriptional addiction. This led to the preferential loss of expression of oncogenes involved in PDGF, EGFR, VEGF, insulin/IGF/MAPKK, FGF, Hedgehog, TGFß, and PI3K signaling pathways. Increased reactive oxygen species level in cancer cells was a prerequisite for targeting the mRNA transcription machinery, thus conferring cancer selectivity to these compounds. Furthermore, DNA repair factors involved in homologous recombination were among the most prominently repressed proteins. In cancer patient samples, RITA, AF, and Onc-1 sensitized to poly(ADP-ribose) polymerase inhibitors both in vitro and ex vivo These findings might pave a way for new synthetic lethal combination therapies.Significance: These findings highlight agents that target transcriptional addiction in cancer cells and suggest combination treatments that target RNA processing and DNA repair pathways simultaneously as effective cancer therapies.

Muscle2View, a CellProfiler pipeline for detection of the capillary-to-muscle fiber interface and high-content quantification of fiber type-specific histology.

Because manual immunohistochemical analysis of features such as skeletal muscle fiber typing, capillaries, myonuclei, and fiber size-related parameters is time consuming and prone to user subjectivity, automatic computational methods could allow for faster and more objective evaluation. Here, we developed Muscle2View, a free CellProfiler-based pipeline that integrates all key fiber-morphological variables, including the novel quantification of the capillary-to-fiber interface, in one single tool. Provided that the images are of sufficient quality and the settings are configured for the specific study, the pipeline allows for automatic and unsupervised analysis of fiber borders, myonuclei, capillaries, and morphometric parameters in a fiber type-specific manner from large batches of images in <10 min/tissue sample. The novel identification of the capillary-to-fiber interface allowed for the calculation of microvascular factors such as capillary contacts (CC), individual capillary-to-fiber ratio (C/Fi), and capillary-to-fiber perimeter exchange (CFPE) index. When comparing the Muscle2View pipeline to manual or semiautomatic analysis, overall the results revealed strong correlations. For several variables, however, there were differences (5-15%) between values computed by manual counting and Muscle2View, suggesting that the methods should not necessarily be used interchangeably. Collectively, we demonstrate that the Muscle2View pipeline can provide unbiased and high-content analysis of muscle cross-sectional immunohistochemistry images. In addition to the classical morphological measurements, the Muscle2View can identify the complex capillary-to-fiber network and myonuclear density in a fiber type-specific manner. This robust analysis is done in one single run within a user-friendly and flexible environment based on the free and widely used image software CellProfiler.NEW & NOTEWORTHY Here, we developed a freely available CellProfiler-based pipeline termed Muscle2View, which provides unbiased, high-content analysis of muscle cross-sectional immunohistochemistry images. In addition to fiber typing, myonuclei counting, and the quantification of fiber type-specific morphological measurements, the Muscle2View pipeline can identify the complex capillary-to-fiber network from a batch of images within minutes. Thus, the Muscle2View is a viable tool for researchers aiming to quantify immunohistochemical variables from skeletal muscle biopsies.

Inhibition of p53 inhibitors: progress, challenges and perspectives.

p53 is the major tumor suppressor and the most frequently inactivated gene in cancer. p53 could be disabled either by mutations or by upstream negative regulators, including, but not limited to MDM2 and MDMX. p53 activity is required for the prevention as well as for the eradication of cancers. Restoration of p53 activity in mouse models leads to the suppression of established tumors of different origin. These findings provide a strong support to the anti-cancer strategy aimed for p53 reactivation. In this review, we summarize recent progress in the development of small molecules, which restore the tumor suppressor function of wild-type p53 and discuss their clinical advance. We discuss different aspects of p53-mediated response, which contribute to suppression of tumors, including non-canonical p53 activities, such as regulation of immune response. While targeting p53 inhibitors is a very promising approach, there are certain limitations and concerns that the intensive research and clinical evaluation of compounds will hopefully help to overcome.

External Validation of the ISAN, A2DS2, and AIS-APS Scores for Predicting Stroke-Associated Pneumonia.

BACKGROUND: The Prestroke Independence, Sex, Age, National Institutes of Health Stroke Scale (ISAN), Age, Atrial Fibrillation, Dysphagia, male sex, and National Institutes of Health Stroke Scale (A2DS2), and acute ischemic stroke-associated pneumonia score (AIS-APS) scores were created to predict stroke-associated pneumonia (SAP), one of the most important medical stroke complications. External validation of all such scores in an acute stroke population was the aim of our study. METHODS: Patients with ischemic or hemorrhagic stroke were prospectively enrolled in the multicenter Stroke-Induced Pneumonia in Andalucía project between October 2014 and May 2016. Receiver operating characteristic curves and linear regression analyses were used to determine discrimination ability of the scores. The Hosmer-Lemeshow goodness-of-fit test and the plot of observed versus predicted SAP risk were used to assess model calibration. RESULTS: Among 201 included patients, SAP rate was 15.5% (31). Higher ISAN, A2DS2, and AIS-APS scores were related to SAP (all P < .001). The C statistic was .83 (95% confidence interval [CI], .76-.91) for the ISAN score, .80 (95% CI, .70-.89) for the A2DS2 score, and .82 (95% CI, .74-.90) for the AIS-APS score, suggesting good discrimination. The ISAN and AIS-APS scores showed good calibration (Cox and Snell R2 = .206 and .174, respectively). The A2DS2 score showed the highest sensitivity (87%), and the AIS-APS score showed the highest specificity (92.8%). CONCLUSIONS: In our cohort, the external validation of ISAN, A2DS2, and AIS-APS scores have demonstrated their accurate prediction of SAP and the ability of these scores as screening tools to better manage SAP. The AIS-APS score would be recommendable for the development of future clinical trials.

Optimization of Hydroxyethylamine Transition State Isosteres as Aspartic Protease Inhibitors by Exploiting Conformational Preferences.

NMR conformational analysis of a hydroxyethylamine peptide isostere developed as an aspartic protease inhibitor shows that it is a flexible architecture. Cyclization to form pyrrolidines, piperidines, or morpholines results in a preorganization of the whole system in solution. The resulting conformation is similar to the conformation of the inhibitor in the active site of BACE-1. This entropic gain results in increased affinity for the enzyme when compared with the acyclic system. For morpholines 27 and 29, the combination of steric and electronic factors is exploited to orient substituents toward S1, S1', and S2' pockets both in the solution and in the bound states. These highly preorganized molecules proved to be the most potent compounds of the series. Additionally, the morpholines, unlike the pyrrolidine and piperidine analogues, have been found to be brain penetrant BACE-1 inhibitors.

TAp73 suppresses tumor angiogenesis through repression of proangiogenic cytokines and HIF-1α activity.

The p53-family member TAp73 is known to function as a tumor suppressor and regulates genomic integrity, cellular proliferation, and apoptosis; however, its role in tumor angiogenesis is poorly understood. Here we demonstrate that TAp73 regulates tumor angiogenesis through repression of proangiogenic and proinflammatory cytokines. Importantly, loss of TAp73 results in highly vascularized tumors, as well as an increase in vessel permeability resulting from disruption of vascular endothelial-cadherin junctions between endothelial cells. In contrast, loss of the oncogenic p73 isoform ΔNp73 leads to reduced blood vessel formation in tumors. Furthermore, we show that up-regulated ΔNp73 levels are associated with increased angiogenesis in human breast cancer and that inhibition of TAp73 results in an accumulation of HIF-1α and up-regulation of HIF-1α target genes. Taken together, our data demonstrate that loss of TAp73 or ΔNp73 up-regulation activates the angiogenic switch that stimulates tumor growth and progression.

Molecular cloning, characterization and gene expression of the full length cDNA encoding the porcine CD11b(αM) and chromosomal localization of the porcine CD11a(αL)-CD11b(αM)-CD11b(αD) gene cluster.

CD11b (α(M)) is a cell-surface glycoprotein mainly expressed on myeloid cells, required for important interactions during the immune response and involved in the pathogenesis of several diseases. The full length cDNA encoding porcine CD11b protein has been cloned and sequenced. Pig CD11b cDNA sequence contains an ORF of 3459 nucleotides that encodes a deduced polypeptide of 1152 amino acid residues that share with CD11b from other species: High % amino acid identity, common domains (α-I, Ca(++) binding, MIDAS), N-glycosylation sites, and the seven FG-GAP tandem repeats. Real time quantitative PCR expression analysis revealed that CD11b transcripts were highly expressed in neutrophils, showing a lower expression in spleen. The CD11a-CD11b-CD11c gene cluster locates on the porcine chromosome region SSC3p15-17.

Hypersomnia as presenting symptom of anti-Ma2-associated encephalitis: case study.

We describe a patient who presented with excessive daytime sleepiness (EDS) and was eventually diagnosed with anti-Ma2 encephalitis. Neurological examination disclosed somnolence, left palpebral ptosis, and vertical gaze paresis. A brain MRI showed high signal intensity in the hypothalamus and each hippocampus. Ma2 antibodies were found in the patient's serum, and fiberbronchoscopy disclosed a lung carcinoma. After three months of steroid treatment, the results of the patient's neurological exam became normal. We conclude that anti-Ma2 encephalitis may present with mostly isolated EDS and that it may respond to steroids despite old age and the presence of an untreated lung cancer.

Molecular cloning, chromosomal location, and expression analysis of porcine CD14.

CD14 is a membrane-associated glycosylphosphatidylinositol (GPI)-anchored protein that binds lipopolysaccharide (LPS) of Gram-negative bacteria and enables LPS-dependent responses in a variety of cells. In this study a cDNA containing the porcine CD14 coding sequence has been cloned and its complete sequence determined. The amino acid sequence deduced from pig CD14 cDNA encodes a 373 amino acid polypeptide that exhibits 75%, 72%, 69%, 66%, 57% and 56% similarity to CD14 from cow, horse, human, rabbit, mouse and rat, respectively. Structural analysis showed that the porcine CD14 is a membrane glycoprotein with a GPI-anchor site and an extracellular domain containing 11 leucine-rich repeats. In addition, the LPS-binding regions identified in the human CD14 are highly conserved in the N-terminal domain of the porcine sequence. Fluorescence in situ hybridization was used to locate the CD14 gene on the pig chromosome 2, band q28. Expression analysis revealed that porcine CD14 transcripts were detected in all tissues and cells examined, suggesting that the expression of porcine CD14 gene is not restricted to myeloid cell lineage. Finally, we report that LPS stimulation significantly up-regulated CD14 gene expression in porcine alveolar macrophages.

Tratamiento implantológico de la zona posterior del maxilar superior. Elevación del seno maxilar / Implant treatment in the posterior maxilla. The maxillary sinus graft

La rehabilitación del sector posterior del maxilar superior con implantes osteointegrados frecuentemente presenta problemas debido a una disponibilidad ósea vertical insuficiente causada por la presencia del seno maxilar. Para solucionar la falta de altura ósea, se hace necesario realizar procedimientos destinados a aumentar la cantidad de hueso en este área, tal como la colocación de injertos óseos dentro del seno, procedimiento conocido como elevación del seno maxilar. El objetivo de esta revisión de la literatura es presentar un análisis detallado de los factores relacionados con esta técnica, desde la anatomía del seno maxilar, la historia, la descripción e indicación de las técnicas, el uso de materiales de relleno, hasta las distintas situaciones que influyen en el éxito del tratamiento y que por lo tanto constituyen factores de riesgo al realizar este procedimiento. (AU)