Miracle-dz/UniFAR-FTUs huggingface dataset:
Circulating Tumor Cell Clusters Are Frequently Detected in Women with Early-Stage Breast Cancer

Simple Summary Metastases cause the majority of breast cancer-related deaths. Circulating tumor cells (CTCs), and in particular CTC-clusters, are considered the seeds of metastasis, but their analysis in the early-stages of the disease has so far been limited by the fact that, by using conventional and epithelial-based technologies (as the FDA-approved CellSearch platform), they are more often detected in the metastatic setting. It is known, however, that cancer cells are heterogeneous and can downregulate the expression of epithelial markers, thus limiting the detection capability of epithelial-based technologies. Here, we show that it is possible to increase CTC-cluster detection by using an epithope-independent technology based on blood filtration, and in particular that this strategy allows to detect a high number of CTC-clusters in stage II-III breast cancer patients, before and during neoadjuvant treatment. Our results therefore offer a new opportunity to deepen our understanding of the cancer dissemination process in its early steps. Abstract The clinical relevance of circulating tumor cell clusters (CTC-clusters) in breast cancer (BC) has been mostly studied using the CellSearch®, a marker-dependent method detecting only epithelial-enriched clusters. However, due to epithelial-to-mesenchymal transition, resorting to marker-independent approaches can improve CTC-cluster detection. Blood samples collected from healthy donors and spiked-in with tumor mammospheres, or from BC patients, were processed for CTC-cluster detection with 3 technologies: CellSearch®, CellSieve™ filters, and ScreenCell® filters. In spiked-in samples, the 3 technologies showed similar recovery capability, whereas, in 19 clinical samples processed in parallel with CellSearch® and CellSieve™ filters, filtration allowed us to detect more CTC-clusters than CellSearch® (median number = 7 versus 1, p = 0.0038). Next, samples from 37 early BC (EBC) and 23 metastatic BC (MBC) patients were processed using ScreenCell® filters for attaining both unbiased enrichment and marker-independent identification (based on cytomorphological criteria). At baseline, CTC-clusters were detected in 70% of EBC cases and in 20% of MBC patients (median number = 2, range 0–20, versus 0, range 0–15, p = 0.0015). Marker-independent approaches for CTC-cluster assessment improve detection and show that CTC-clusters are more frequent in EBC than in MBC patients, a novel finding suggesting that dissemination of CTC-clusters is an early event in BC natural history.
Vibrio Species in an Urban Tropical Estuary: Antimicrobial Susceptibility, Interaction with Environmental Parameters, and Possible Public Health Outcomes

The genus Vibrio comprises pathogens ubiquitous to marine environments. This study evaluated the cultivable Vibrio community in the Guanabara Bay (GB), a recreational, yet heavily polluted estuary in Rio de Janeiro, Brazil. Over one year, 66 water samples from three locations along a pollution gradient were investigated. Isolates were identified by MALDI-TOF mass spectrometry, revealing 20 Vibrio species, including several potential pathogens. Antimicrobial susceptibility testing confirmed resistance to aminoglycosides, beta-lactams (including carbapenems and third-generation cephalosporins), fluoroquinolones, sulfonamides, and tetracyclines. Four strains were producers of extended-spectrum beta-lactamases (ESBL), all of which carried beta-lactam and heavy metal resistance genes. The toxR gene was detected in all V. parahaemolyticus strains, although none carried the tdh or trh genes. Higher bacterial isolation rates occurred in months marked by higher water temperatures, lower salinities, and lower phosphorus and nitrogen concentrations. The presence of non-susceptible Vibrio spp. was related to indicators of eutrophication and sewage inflow. DNA fingerprinting analyses revealed that V. harveyi and V. parahaemolyticus strains non-susceptible to antimicrobials might persist in these waters throughout the year. Our findings indicate the presence of antimicrobial-resistant and potentially pathogenic Vibrio spp. in a recreational environment, raising concerns about the possible risks of human exposure to these waters.
Biochemical Validation of a Self-Administered Food Frequency Questionnaire to Assess Diet Using Carotenoids and Vitamins E and D in Male Adolescents in Spain

Reliable tools to evaluate diet are needed, particularly in life periods such as adolescence in which a rapid rate of growth and development occurs. We assessed the biochemical validity of a self-administered food frequency questionnaire (FFQ) in a sample of Spanish male adolescents using carotenoids and vitamin E and D data. We analyzed data from 122 male adolescents aged 15–17 years of the INMA-Granada birth cohort study. Adolescents answered a 104-item FFQ and provided a non-fasting blood sample. Mean daily nutrient intakes and serum concentration were estimated for main carotenoids (lutein-zeaxanthin, β-cryptoxanthin, lycopene, α-carotene and β-carotene), vitamins E and D and also for fruit and vegetable intake. Pearson correlation coefficients (r) and the percentage of agreement (same or adjacent quintiles) between serum vitamin concentrations and energy-adjusted intakes were estimated. Statistically significant correlation coefficients were observed for the total carotenoids (r = 0.40) and specific carotenoids, with the highest correlation observed for lutein–zeaxanthin (r = 0.42) and the lowest for β-carotene (0.23). The correlation coefficient between fruit and vegetable intake and serum carotenoids was 0.29 (higher for vegetable intake, r = 0.33 than for fruit intake, r = 0.19). Low correlations were observed for vitamin E and D. The average percentage of agreement for carotenoids was 55.8%, and lower for vitamin E and D (50% and 41%, respectively). The FFQ may be an acceptable tool for dietary assessment among male adolescents in Spain.
The Lived Experiences of Stigmatization in the Process of HIV Status Disclosure among People Living with HIV in Taiwan

People living with HIV (PLWH) face social stigma which makes disclosure of HIV status difficult. The purpose of this descriptive qualitative study was to understand the lived experiences of stigmatization in the process of disease disclosure among PLWH in Taiwan. Analysis of the semi-structured interviews from 19 PLWH in Taiwan revealed two phases and six themes. Phase one “experiences before disclosure” involved three themes: “Struggles under the pressure of concealing the HIV Status”, “Torn between fear of unemployment/isolation and desire to protect closed ones”, and “Being forced to disclose the HIV status. ” Phase two “experiences after disclosure” included three themes: “Receiving special considerations and requirements from school or work”, “Receiving differential treatments in life and when seeking medical care”, and “Stress relief and restart. ” Healthcare professionals need to assess stigmatization in PLWH and develop individualized approaches to assist with the disease disclosure process.
Synthetic Studies on Isoprenoidquinones. I. A Facile Regioand Stereocontrolled Synthesis of Protected Hydroquinones with an Omega-Hydroxyprenyl or Omega-Hydroxygeranyl Side Chain

Regio-and stereocontrolled terminal functionalization of protected 2-prenyl-and 2-geranylhydroquinones (2) was achieved by way of terminal methallylic sulfides (4), which were easily obtained via benzenesulfenyl chloride addition followed by dehydrochlorination or via allylic chlorination with SO2Cl2 followed by sulfenylation, leading to terminal trans-allylic alcohols (5) which are promising intermediates for the synthesis of polyisoprenoidquinones. In this transformation, interesting steric effects of the hydroquinone nucleus with the adjacent arylic methyl group influencing the site-selectivity in the reactions of the isoprenoid side chain were observed.
Cymbopogon winterianus Essential Oil Attenuates Bleomycin-Induced Pulmonary Fibrosis in a Murine Model

The essential oil of Cymbopogon winterianus (EOCW) is a natural product with antioxidant, anti-inflammatory, and antifibrotic properties. We studied the effect of EOCW in the progression of histological changes of pulmonary fibrosis (PF) in a rodent model. The oil was obtained by hydrodistillation and characterized using gas chromatography–mass spectrometry. Intratracheal instillation of bleomycin was performed in 30 rats to induce PF, while Sham animals were subjected to instillation of saline solution. The treatment was performed using daily oral administration of distilled water, EOCW at 50, 100, and 200 mg/kg, and deflazacort (DFC). After 28 days, hemogram and bronchoalveolar lavage fluid (BALF), tissue levels of malondialdehyde (MDA), superoxide dismutase (SOD), and catalase (CAT) were assayed. Histological grading of PF, immunohistochemical expression of α-smooth muscle actin (α-SMA), and transforming growth factor-β (TGF-β) were also analyzed. The EOCW major compounds were found to be citronellal, geraniol, and citronellol. EOCW significantly reduced inflammation in BALF, reduced MDA levels, and increased SOD activity. EOCW attenuated histological grading of PF and reduced immunohistochemical expression of α-SMA and TGF-β in a dose-dependent way, likely due to the reduction of oxidative stress, inflammation, and TGF-β-induced myofibroblast differentiation.
Effect of the COVID-19 Pandemic on Medical Student Career Perceptions: Perspectives from Medical Students in China

Our aim was to examine perceived occupational turnover intentions among medical students and the associated factors. A cross-sectional study using a Web-based survey was conducted. A total of 2922 completed responses were received (response rate 55.7%). A total of 58.4% (95% CI 56.6–60.2) reported high turnover intention (score of 7–15). The odds of higher total turnover score among the fifth-year students was nearly four times that of first-year students (OR = 3.88, 95% CI 2.62–5.73). Perception of the medical profession as not being of high social status and reputation significantly influenced high turnover intention scores (OR = 2.26, 95% CI 1.90–2.68). All three dimensions of the multidimensional scale of perceived social support (MSPSS) significantly predict turnover intention. Lower scores in the support from Significant Other (OR = 1.47, 95% CI 1.17–1.84), Family (OR = 1.47, 95% CI 1.18–1.83) and Friend (OR = 1.42, 95% CI 1.14–1.77) subscales were associated with higher turnover intention. Low score in the Brief Resilience Scale (BRS) was also associated with higher turnover intention (OR = 1.44, 95% CI 1.17–1.77). The findings shed light on the importance of changing public attitudes towards respecting the medical profession and improving the implementation of policies to protect the well-being of people in the medical profession.
Disparities in the Evolution of the COVID-19 Pandemic between Spanish Provinces

Spain experienced a second wave of the COVID-19 pandemic in autumn 2020, which has been approached with different measures by regional authorities. We analyze the presence of convergence in the cumulative incidence for 14 days (CI14) in provinces and self-governing cities. The Phillips–Sul methodology was used to study the grouping of behavior between provinces, and an ordered logit model was estimated to understand the forces that drive creating the different convergence clubs. We reject the presence of a single pattern of behavior in the evolution of the CI14 across territories. Four statistically different convergence clubs and an additional province (Madrid) with divergent behavior are observed. Provinces with developed agricultural and industrial economic sectors, high mobility, and a high proportion of Central and South American immigrants had the highest level of CI14. We show that the transmission of the virus is not homogeneous in the Spanish national territory. Our results are helpful for identifying differences in determinants that could explain the pandemic’s evolution and for formulating hypotheses about the effectiveness of implemented measures.
Chronic low back pain, Modic changes and low-grade virulent infection: efficacy of antibiotic treatment

Chronic low back pain (CLBP) has consistently been associated with the longest number of years lived with a disability in global studies, while commonly used treatments for CLBP are largely ineffective. In 2013 a randomized, double-blind, controlled study demonstrated significant improvements in CLBP patients demonstrating Modic changes type 1 on their MRI scans and undergoing long-term oral antibiotic treatment (100 days). Much of the ensuing debate has focused on whether this was a true infection or contamination. Newer and more advanced technologies clearly point to an ongoing low-grade infection. We have reviewed all of the clinical trials published in the recent past and conclude that there is compelling evidence for the effect of long-term oral antibiotic treatment for this patient group.
Exogenous Potassium (K+) Positively Regulates Na+/H+ Antiport System, Carbohydrate Metabolism, and Ascorbate–Glutathione Cycle in H2S-Dependent Manner in NaCl-Stressed Tomato Seedling Roots

Potassium (K+) is one of the vital macronutrients required by plants for proper growth and blossoming harvest. In addition, K+ also plays a decisive role in promoting tolerance to various stresses. Under stressful conditions, plants deploy their defense system through various signaling molecules, including hydrogen sulfide (H2S). The present investigation was carried out to unravel the role of K+ and H2S in plants under NaCl stress. The results of the study show that NaCl stress caused a reduction in K+ and an increase in Na+ content in the tomato seedling roots which coincided with a lower H+-ATPase activity and K+/Na+ ratio. However, application of 5 mM K+, in association with endogenous H2S, positively regulated the Na+/H+ antiport system that accelerated K+ influx and Na+ efflux, resulting in the maintenance of a higher K+/Na+ ratio. The role of K+ and H2S in the regulation of the Na+/H+ antiport system was validated by applying sodium orthovanadate (plasma membrane H+-ATPase inhibitor), tetraethylammonium chloride (K+ channel blocker), amiloride (Na+/H+ antiporter inhibitor), and hypotaurine (HT, H2S scavenger). Application of 5 mM K+ positively regulated the ascorbate–glutathione cycle and activity of antioxidant enzymes that resulted in a reduction in reactive oxygen species generation and associated damage. Under NaCl stress, K+ also activated carbohydrate metabolism and proline accumulation that caused improvement in osmotic tolerance and enhanced the hydration level of the stressed seedlings. However, inclusion of the H2S scavenger HT reversed the effect of K+, suggesting H2S-dependent functioning of K+ under NaCl stress. Therefore, the present findings report that K+, in association with H2S, alleviates NaCl-induced impairments by regulating the Na+/H+ antiport system, carbohydrate metabolism, and antioxidative defense system.
Survival Prediction in Intrahepatic Cholangiocarcinoma: A Proof of Concept Study Using Artificial Intelligence for Risk Assessment

Several scoring systems have been devised to objectively predict survival for patients with intrahepatic cholangiocellular carcinoma (ICC) and support treatment stratification, but they have failed external validation. The aim of the present study was to improve prognostication using an artificial intelligence-based approach. We retrospectively identified 417 patients with ICC who were referred to our tertiary care center between 1997 and 2018. Of these, 293 met the inclusion criteria. Established risk factors served as input nodes for an artificial neural network (ANN). We compared the performance of the trained model to the most widely used conventional scoring system, the Fudan score. Predicting 1-year survival, the ANN reached an area under the ROC curve (AUC) of 0.89 for the training set and 0.80 for the validation set. The AUC of the Fudan score was significantly lower in the validation set (0.77, p < 0.001). In the training set, the Fudan score yielded a lower AUC (0.74) without reaching significance (p = 0.24). Thus, ANNs incorporating a multitude of known risk factors can outperform conventional risk scores, which typically consist of a limited number of parameters. In the future, such artificial intelligence-based approaches have the potential to improve treatment stratification when models trained on large multicenter data are openly available.
Determinants of Satisfaction with Services, and Trust in the Information Received in Community Pharmacies: A Comparative Analysis to Foster Pharmaceutical Care Adoption

Patient’s satisfaction with community pharmacy services, and patients’ trust in the information received in community pharmacies are important drivers of pharmaceutical care adoption. An online questionnaire assessing patient satisfaction with the services received in pharmacies and trust in the pharmacist’s advice, along with their determinants, was administered to 343 Romanian chronic and non-chronic patients. Using various statistical tests, exploratory factor analysis, and robust regression we explored determinants of satisfaction and trust. We found that satisfaction with services is predicted by pharmacists’ attitude (β = 631, p < 0.001), low waiting time (β = 0.180, p < 0.001), affordable cost of the drugs (β = 0.09, p = 0.009), and drug availability (β = 0.157, p < 0.001). At the same time, trust in the information received is driven by pharmacists’ attention (β = 0.610, p < 0.001), whether the patient received precautionary information (β = 0.425, p < 0.001), low waiting time (β = 0.287, p < 0.001), and whether the respondent is a chronic patient or not (non-chronic patients express more trust, β = 0.328, p = 0.04). Our study expands the existing paradigm that sees trust as a simple predictor of satisfaction by showing that trust and satisfaction are predicted by different variables, and thus they should be addressed using different strategies. In fact, we found that they share only one predictor—waiting time, highly significant in both cases. Our findings show that, without prioritizing trust in the information received in community pharmacies to reduce information asymmetry between patient and pharmacist, the focus only on patient satisfaction may lead to a scenario in which community pharmacies will end up to be better integrated in the business sector and not in the public health system.
Mutation in a SARS-CoV-2 Haplotype from Sub-Antarctic Chile Reveals New Insights into the Spike’s Dynamics

The emergence of SARS-CoV-2 variants, as observed with the D614G spike protein mutant and, more recently, with B.1. 1.7 (501Y.V1), B.1. 351 (501Y.V2) and B.1. 1.28. 1 (P.1) lineages, represent a continuous threat and might lead to strains of higher infectivity and/or virulence. We report on the occurrence of a SARS-CoV-2 haplotype with nine mutations including D614G/T307I double-mutation of the spike. This variant expanded and completely replaced previous lineages within a short period in the subantarctic Magallanes Region, southern Chile. The rapid lineage shift was accompanied by a significant increase of cases, resulting in one of the highest incidence rates worldwide. Comparative coarse-grained molecular dynamic simulations indicated that T307I and D614G belong to a previously unrecognized dynamic domain, interfering with the mobility of the receptor binding domain of the spike. The T307I mutation showed a synergistic effect with the D614G. Continuous surveillance of new mutations and molecular analyses of such variations are important tools to understand the molecular mechanisms defining infectivity and virulence of current and future SARS-CoV-2 strains.
Expression Analysis of FGF/FGFR and FOX Family Proteins in Mucosal Tissue Obtained from Orofacial Cleft-Affected Children

Simple Summary Craniofacial development is an intricate and delicate process in normal embryogenesis requiring spatiotemporal release of various mediators/proteins that provide communication between different cell/tissue types, like epithelial cells, connective tissue, and endothelial cells. If this sequence is impaired or blocked due to genetic or environmental factors, it can lead to clefting. A cleft is an abnormal space or gap in the upper lip, alveolus, or palate that occurs due to failure of completion of fusion processes in the midline during facial development. Previous studies have identified various genetic factors (genes) that can lead to clefting. The most promising candidates amongst them are FGF/FGFR (fibroblast growth factor/FGF receptor) signaling genes and FOX (forkhead box protein) genes. We investigated the expression of these genes in tissue material obtained from cleft-affected patients. Our results indicate that these genes profoundly affect the pathogenesis and manifestation of clefts, especially by enhancing local site inflammation and fibrosis. Further, they play a vital role in angiogenesis, apoptosis, and cell proliferation. Abstract Orofacial clefts affect hundreds of thousands of children worldwide annually and are usually corrected by a series of surgeries extending to childhood. The underlying mechanisms that lead to clefts are still unknown, mainly because of the multifactorial etiology and the myriad of interactions between genes and environmental factors. In the present study, we investigated the role and expression of candidate genes belonging to the FGF/FGFR signaling pathway and FOX family in tissue material obtained from 12 pediatric patients undergoing cleft correction surgery. The expression was investigated using immunohistochemistry (IHC) and chromogenic in-situ hybridization (CISH) in three cell/tissue types—epithelial cells, connective tissue, and endothelial cells. We found elevated expression of FGFR1 in epithelial cells while no expression was observed in endothelial cells. Further, our results elucidate the potential pathogenetic role of FGFR1 in cellular proliferation, local site inflammation, and fibrosis in cleft patients. Along with bFGF (also called FGF2), FGFR1 could play a pro-inflammatory role in clefts. Over-amplification of FGFR2 in some patients, along with bFGF, could potentially suggest roles for these genes in angiogenesis. Additionally, increased expression of FOXE1 (also called TTF2) contributes to local site inflammation. Finally, zero to low amplification of FOXO1 could suggest its potential role in inducing oxidative stress in the endothelium along with reduced epithelial apoptosis.