Miracle-dz/UniFAR-FTUs huggingface dataset:
A Fluorescent Real-Time Plaque Assay Enables Single-Cell Analysis of Virus-Induced Cytopathic Effect by Live-Cell Imaging

Conventional plaque assays rely on the use of overlays to restrict viral infection allowing the formation of distinct foci that grow in time as the replication cycle continues leading to countable plaques that are visualized with standard techniques such as crystal violet, neutral red, or immunolabeling. This classical approach takes several days until large enough plaques can be visualized and counted with some variation due to subjectivity in plaque recognition. Since plaques are clonal lesions produced by virus-induced cytopathic effect, we applied DNA fluorescent dyes with differential cell permeability to visualize them by live-cell imaging. We could observe different stages of that cytopathic effect corresponding to an early wave of cells with chromatin-condensation followed by a wave of dead cells with membrane permeabilization within plaques generated by different animal viruses. This approach enables an automated plaque identification using image analysis to increase single plaque resolution compared to crystal violet counterstaining and allows its application to plaque tracking and plaque reduction assays to test compounds for both antiviral and cytotoxic activities. This fluorescent real-time plaque assay sums to those next-generation technologies by combining this robust classical method with modern fluorescence microscopy and image analysis approaches for future applications in virology.
The Association between Parental Support and Adolescents’ Psychological Complaints: The Mediating Role of a Good School Climate

Parental support is an important factor affecting young people’s mental well-being, but the school climate also plays an important role. However, few studies have previously examined whether the school climate serves as a mediator for adolescents’ mental health problems. This study aimed to investigate the association between parental support and students’ psychological complaints, while also examining the possible mediating role that a good school climate may have. Data derives from 5783 senior-level students (age 15–16) distributed over 152 school units in Stockholm municipality. Regression linear analysis was used for the analysis and Baron and Kenny’s four-step mediation model has been applied. Sobel’s test was conducted in order to test the significance of the mediation effect. The results show that there is a significant negative association between parental support and students’ psychological complaints, and that school climate has a mediating role in this association. It can be concluded that school climate has a partly mediating role in the association between parental support and students’ psychological complaints. Therefore, it seems important to develop the school climate in order to strengthen this source of support to reduce mental health problems among adolescents.
Establishment of a Pretreatment Nomogram to Predict the 6-Month Mortality Rate of Patients with Advanced Biliary Tract Cancers Undergoing Gemcitabine-Based Chemotherapy

Simple Summary The development of a simple tool that uses pretreatment clinical factors to predict the 6-month mortality rate of patients with advanced biliary tract cancer is critical in order to assist physicians in evaluating treatment options and outcomes. We established a nomogram including four independent pretreatment factors—gender, monocyte to lymphocyte ratio, alkaline phosphatase, and liver metastasis—based on data from 202 patients undergoing gemcitabine-based chemotherapy. The performance of this nomogram for 6-month mortality-risk prediction was promising and feasible, providing clinicians and patients with additional information for evaluating therapeutic options. Abstract Background: The estimation of mortality risk among patients diagnosed with advanced cancer provides important information for clinicians and patients in clinical practice. Currently, gemcitabine-based chemotherapy regimens are the standard treatment for patients with advanced biliary tract cancer (BTC). We aimed to develop a nomogram to predict the 6-month mortality rate among patients with advanced BTC to help physicians evaluate treatment options and outcomes. Patients: We conducted a retrospective analysis to evaluate the 6-month mortality rate among patients with advanced BTC who underwent gemcitabine-based chemotherapy from 2012 to 2018. Data regarding pretreatment factors and the clinical response to treatment were collected. Univariate and multivariate analyses were performed to identify independent factors for nomogram creation. Results: A total of 202 advanced BTC patients who were treated with gemcitabine-based chemotherapy were included in this analysis. No difference in survival was identified between patients undergoing gemcitabine monotherapy and those treated with gemcitabine combined with other cytotoxic agents. The univariate analysis revealed 10 significant factors, while the multivariate analysis identified four independent factors, including gender, monocyte to lymphocyte ratio (MLR), alkaline phosphatase (ALP), and liver metastasis, which were used to establish the nomogram. The performance of this nomogram for the prediction of 6-month mortality risk was found to be promising and feasible based on logistic regression. Conclusion: A nomogram based on four independent pretreatment factors, including gender, MLR, ALP, and liver metastasis, was established to predict the 6-month mortality risk in patients with advanced BTC; it can provide clinicians and patients with additional information when evaluating treatment outcomes.
Medical Cost of Cancer Care for Privately Insured Children in Chile

Medical care for children with cancer is complex and expensive, and represents a large financial burden for families around the world. We estimated the medical cost of cancer care for children under the age of 18, using administrative records of the universe of children with private insurance in Chile in the period 2007–2018, based on a sample of 3853 observations. We analyzed total cost and out-of-pocket spending by patients’ characteristics, type of cancer, and by service. Children with cancer had high annual medical costs, USD 32,287 on average for 2018. Costs were higher for the younger children in the sample. The vast majority of the cost was driven by inpatient hospital care for all types of cancer. The average total cost increased 20% in real terms over the period of study, while out-of-pocket expenses increased almost 29%. Private insurance beneficiaries faced a significant economic burden associated with medical treatment of a child with cancer. Interventions that reduce hospitalizations, as well as systemwide reforms that incorporate maximum out-of-pocket payments and prevent catastrophic expenditures, can contribute to alleviating the financial burden of childhood cancer.
Perinatal High-Salt Diet Induces Gut Microbiota Dysbiosis, Bile Acid Homeostasis Disbalance, and NAFLD in Weanling Mice Offspring

A perinatal high-salt (HS) diet was reported to elevate plasma triglycerides. This study aimed to investigate the hypothesis that a perinatal HS diet predisposed offspring to non-alcoholic fatty liver disease (NAFLD), the hepatic manifestation of abnormal lipid metabolism, and the possible mechanism. Female C57BL/6 mice were fed a control diet (0.5% NaCl) or HS diet (4% NaCl) during pregnancy and lactation and their offspring were sacrificed at weaning. The perinatal HS diet induced greater variation in fecal microbial beta-diversity (β-diversity) and increased bacteria abundance of Proteobacteria and Bacteroides. The gut microbiota dysbiosis promoted bile acid homeostasis disbalance, characterized by the accumulation of lithocholic acid (LCA) and deoxycholic acid (DCA) in feces. These alterations disturbed gut barrier by increasing the expression of tight junction protein (Tjp) and occludin (Ocln), and increased systemic lipopolysaccharide (LPS) levels and hepatic inflammatory cytokine secretion (TNF-α and IL-6) in the liver. The perinatal HS diet also inhibited hepatic expression of hepatic FXR signaling (CYP7A1 and FXR), thus triggering increased hepatic expression of pro-inflammatory cytokines (TNF-α and IL-6) and hepatic lipid metabolism-associated genes (SREBP-1c, FAS, ACC), leading to unique characteristics of NAFLD. In conclusion, a perinatal HS diet induced NAFLD in weanling mice offspring; the possible mechanism was related to increased bacteria abundance of Proteobacteria and Bacteroides, increased levels of LCA and DCA in feces, and increased expressions of hepatic FXR signaling.
Oropharyngeal Squamous Cell Carcinoma Treatment in the Era of Immune Checkpoint Inhibitors

While head and neck squamous cell carcinomas (HNSCC) are marginally decreasing due to the reduction in exposure to the major risk factors, tobacco and alcohol, the incidence of high-risk human papillomavirus (HPV)-positive oropharynx squamous cell carcinomas (OPSCC), especially those in the tonsil and base of tongue subsites, are increasing. Patients with the latter are younger, display a longer overall survival, and show a lower recurrence rate after standard-of-care treatment than those with HPV-negative OPSCC. This may reflect an important role for immune surveillance and control during the natural history of the virally driven tumour development. Immune deviation through acquisition of immune-suppressive factors in the tumour microenvironment (TME) is discussed in relation to treatment response. Understanding how the different immune factors are integrated in the TME battleground offers opportunities for identifying prognostic biomarkers as well as novel therapeutic strategies. OPSCC generally receive surgery or radiotherapy for early-stage tumour treatment, but many patients present with locoregionally advanced disease requiring multimodality therapies which can involve considerable complications. This review focuses on the utilization of newly emerged immune checkpoint inhibitors (PD-1/PD-L1 pathway) for treatment of HNSCC, in particular HPV-positive OPSCC, since they could be less toxic and more efficacious. PD-1/PD-L1 expression in the TME has been extensively investigated as a biomarker of patient response but is yet to provide a really effective means for stratification of treatment. Extensive testing of combinations of therapeutic approaches by types and sequencing will fuel the next evolution of treatment for OPSCC.
Dedicated Algorithm for Unobtrusive Fetal Heart Rate Monitoring Using Multiple Dry Electrodes

Multi-channel measurements from the maternal abdomen acquired by means of dry electrodes can be employed to promote long-term monitoring of fetal heart rate (fHR). The signals acquired with this type of electrode have a lower signal-to-noise ratio and different artifacts compared to signals acquired with conventional wet electrodes. Therefore, starting from the benchmark algorithm with the best performance for fHR estimation proposed by Varanini et al., we propose a new method specifically designed to remove artifacts typical of dry-electrode recordings. To test the algorithm, experimental textile electrodes were employed that produce artifacts typical of dry and capacitive electrodes. The proposed solution is based on a hybrid (hardware and software) pre-processing step designed specifically to remove the disturbing component typical of signals acquired with these electrodes (triboelectricity artifacts and amplitude modulations). The following main processing steps consist of the removal of the maternal ECG by blind source separation, the enhancement of the fetal ECG and identification of the fetal QRS complexes. Main processing is designed to be robust to the high-amplitude motion artifacts that corrupt the acquisition. The obtained denoising system was compared with the benchmark algorithm both on semi-simulated and on real data. The performance, quantified by means of sensitivity, F1-score and root-mean-square error metrics, outperforms the performance obtained with the original method available in the literature. This result proves that the design of a dedicated processing system based on the signal characteristics is necessary for reliable and accurate estimation of the fHR using dry, textile electrodes.
A Transcriptomic Approach to the Metabolism of Tetrapyrrolic Photosensitizers in a Marine Annelid

The past decade has seen growing interest in marine natural pigments for biotechnological applications. One of the most abundant classes of biological pigments is the tetrapyrroles, which are prized targets due their photodynamic properties; porphyrins are the best known examples of this group. Many animal porphyrinoids and other tetrapyrroles are produced through heme metabolic pathways, the best known of which are the bile pigments biliverdin and bilirubin. Eulalia is a marine Polychaeta characterized by its bright green coloration resulting from a remarkably wide range of greenish and yellowish tetrapyrroles, some of which have promising photodynamic properties. The present study combined metabolomics based on HPLC-DAD with RNA-seq transcriptomics to investigate the molecular pathways of porphyrinoid metabolism by comparing the worm’s proboscis and epidermis, which display distinct pigmentation patterns. The results showed that pigments are endogenous and seemingly heme-derived. The worm possesses homologs in both organs for genes encoding enzymes involved in heme metabolism such as ALAD, FECH, UROS, and PPOX. However, the findings also indicate that variants of the canonical enzymes of the heme biosynthesis pathway can be species- and organ-specific. These differences between molecular networks contribute to explain not only the differential pigmentation patterns between organs, but also the worm’s variety of novel endogenous tetrapyrrolic compounds.
Genomic Analysis of Prophages Recovered from Listeria monocytogenes Lysogens Found in Seafood and Seafood-Related Environment

A prophage is a phage-related sequence that is integrated into a bacterial chromosome. Prophages play an important role in bacterial evolution, survival, and persistence. To understand the impact of Listeria prophages on their host genome organizations, this work sequenced two L. monocytogenes strains (134LM and 036LM), previously identified as lysogens by mitomycin C induction. Draft genomes were generated with assembly sizes of 2,953,877 bp and 3,000,399 bp. One intact prophage (39,532 bp) was inserted into the comK gene of the 134LM genome. Two intact prophages (48,684 bp and 39,488 bp) were inserted in tRNA-Lys and elongation-factor genes of the 036LM genome. The findings confirmed the presence of three corresponding induced phages previously obtained by mitomycin C induction. Comparative genomic analysis of three prophages obtained in the newly sequenced lysogens with 61 prophages found in L. monocytogenes genomes, available in public databases, identified six major clusters using whole genome-based phylogenetic analysis. The results of the comparative genomic analysis of the prophage sequences provides knowledge about the diversity of Listeria prophages and their distribution among Listeria genomes in diverse environments, including different sources or geographical regions. In addition, the prophage sequences and their insertion sites contribute to the genomic diversity of L. monocytogenes genomes. These data of prophage sequences, prophage insertion sites, and prophage sequence comparisons, together with ANIb confirmation, could be useful for L. monocytogenes classification by prophages. One potential development could be refinement of prophage typing tools for monitoring or surveillance of L. monocytogenes contamination and transmission.
Biphasic α2β1 Integrin Expression in Breast Cancer Metastasis to Bone

Integrins participate in the pathogenesis and progression of tumors at many stages during the metastatic cascade. However, current evidence for the role of integrins in breast cancer progression is contradictory and seems to be dependent on tumor stage, differentiation status, and microenvironmental influences. While some studies suggest that loss of α2β1 enhances cancer metastasis, other studies suggest that this integrin is pro-tumorigenic. However, few studies have looked at α2β1 in the context of bone metastasis. In this study, we aimed to understand the role of α2β1 integrin in breast cancer metastasis to bone. To address this, we utilized in vivo models of breast cancer metastasis to bone using MDA-MB-231 cells transfected with an α2 expression plasmid (MDA-OEα2). MDA cells overexpressing the α2 integrin subunit had increased primary tumor growth and dissemination to bone but had no change in tumor establishment and bone destruction. Further in vitro analysis revealed that tumors in the bone have decreased α2β1 expression and increased osteolytic signaling compared to primary tumors. Taken together, these data suggest an inverse correlation between α2β1 expression and bone-metastatic potential. Inhibiting α2β1 expression may be beneficial to limit the expansion of primary tumors but could be harmful once tumors have established in bone.
The Longitudinal Link between Organizational Citizenship Behaviors and Three Different Models of Happiness

A growing body of research conducted in general life settings has found positive associations between happiness and prosocial behavior. Unfortunately, equivalent studies in the workplace are lacking. Organizational citizenship behaviors (OCBs), the prosocial behaviors at work, have not been properly studied in relation to happiness, despite the positive consequences of both constructs for workers and companies. In response, our research aims to better understand this relationship from several angles. First, using a three-wave longitudinal design, we explored how OCBs and happiness are related to each other over time. Second, happiness was measured from a broad perspective, and three conceptualizations were adopted: the hedonic (e.g., positive affect and life satisfaction), the eudaimonic (e.g., relatedness and autonomy), and the flourishing (e.g., meaning and engagement) approaches. Thus, not only the prospective link between OCBs and happiness was tested, but it was also explored using the three models of happiness previously mentioned. Third, we conducted this longitudinal design in a less typical sample than previous research (i.e., Chile). We found results that supported our main hypotheses: (1) OCBs are prospective positive predictors of hedonic happiness, eudaimonic happiness, and flourishing; (2) the three models of happiness also prospectively predict OCBs. Our findings suggest that OCBs foster a broad range of happiness facets, which in turn fosters back the emergence of more OCBs, leading to a virtuous circle of prosociality and well-being in the workplace. This positive spiral benefits not only workers’ quality of life, but also organizations’ profitability and sustainability. Theoretical and applied implications for the field of Positive Organizational Psychology are discussed.
Changes in the structure and composition of the ‘Mexical’ scrubland bee community along an elevational gradient

‘Mexical’ scrubland is a sclerophyllous evergreen Mediterranean-like vegetation occurring in the leeward slopes of the main Mexican mountain ranges, under tropical climate. This biome occupies an elevational range approximately from 1900 to 2600 meters above sea level, which frequently is the upper-most part of the mountains range. This puts it at risk of extinction in a scenario of global warming in which an upward retraction of this type of vegetation is expected. The Mexical remains one of the least studied ecosystems in Mexico. For instance, nothing is known about pollinator fauna of this vegetation. Our main objective is to make a first insight into the taxonomic identity of the bee fauna that inhabits this biome, and to study how it is distributed along the elevational gradient that it occupies. Our results highlight that elevation gradient negatively affects bee species richness and that this relationship is strongly mediated by temperature. Bee abundance had no significant pattern along elevational gradient, but shows a significant relationship with flower density. Interestingly, and contrary to previous works, we obtained a different pattern for bee richness and bee abundance. Bee community composition changed strongly along elevation gradient, mainly in relation to temperature and flower density. In a global warming scenario, as temperatures increases, species with cold preferences, occupying the highest part of the elevation gradient, are likely to suffer negative consequences (even extinction risk), if they are not flexible enough to adjust their physiology and/or some life-story traits to warmer conditions. Species occupying mid and lower elevations are likely to extend their range of elevational distribution towards higher ranges. This will foreseeably cause a new composition of species and a new scenario of interactions, the adjustment of which still leaves many unknowns to solve.
Discovery of New Pyrazolopyridine, Furopyridine, and Pyridine Derivatives as CDK2 Inhibitors: Design, Synthesis, Docking Studies, and Anti-Proliferative Activity

New pyridine, pyrazoloyridine, and furopyridine derivatives substituted with naphthyl and thienyl moieties were designed and synthesized starting from 6-(naphthalen-2-yl)-2-oxo-4-(thiophen-2-yl)-1,2-dihydropyridine-3-carbonitrile (1). The chloro, methoxy, cholroacetoxy, imidazolyl, azide, and arylamino derivatives were prepared to obtain the pyridine-−C2 functionalized derivatives. The derived pyrazolpyridine-N-glycosides were synthesized via heterocyclization of the C2-thioxopyridine derivative followed by glycosylation using glucose and galactose. The furopyridine derivative 14 and the tricyclic pyrido[3′,2′:4,5]furo[3,2-d]pyrimidine 15 were prepared via heterocyclization of the ester derivative followed by a reaction with formamide. The newly synthesized compounds were evaluated for their ability to in vitro inhibit the CDK2 enzyme. In addition, the cytotoxicity of the compounds was tested against four different human cancer cell lines (HCT-116, MCF-7, HepG2, and A549). The CDK2/cyclin A2 enzyme inhibitory results revealed that pyridone 1, 2-chloro-6-(naphthalen-2-yl)-4-(thiophen-2-yl)nicotinonitrile (4), 6-(naphthalen-2-yl)-4-(thiophen-2-yl)-1H-pyrazolo[3,4-b]pyridin-3-amine (8), S-(3-cyano-6-(naphthaen-2-yl)-4-(thiophen-2-yl)pyridin-2-yl) 2-chloroethanethioate (11), and ethyl 3-amino-6-(naphthalen-2-yl)-4-(thiophen-2-yl)furo[2,3-b]pyridine-2-carboxylate (14) are among the most active inhibitors with IC50 values of 0.57, 0.24, 0.65, 0.50, and 0.93 µM, respectively, compared to roscovitine (IC50 0.394 μM). Most compounds showed significant inhibition on different human cancer cell lines (HCT-116, MCF-7, HepG2, and A549) with IC50 ranges of 31.3–49.0, 19.3–55.5, 22.7–44.8, and 36.8–70.7 μM, respectively compared to doxorubicin (IC50 40.0, 64.8, 24.7 and 58.1 µM, respectively). Furthermore, a molecular docking study suggests that most of the target compounds have a similar binding mode as a reference compound in the active site of the CDK2 enzyme. The structural requirements controlling the CDK2 inhibitory activity were determined through the generation of a statistically significant 2D-QSAR model.