Miracle-dz/UniFAR-FTUs huggingface dataset:
From Light Microscopy to Analytical Scanning Electron Microscopy (SEM) and Focused Ion Beam (FIB)/SEM in Biology: Fixed Coordinates, Flat Embedding, Absolute References

Abstract Correlative light and electron microscopy (CLEM) has been in use for several years, however it has remained a costly method with difficult sample preparation. Here, we report a series of technical improvements developed for precise and cost-effective correlative light and scanning electron microscopy (SEM) and focused ion beam (FIB)/SEM microscopy of single cells, as well as large tissue sections. Customized coordinate systems for both slides and coverslips were established for thin and ultra-thin embedding of a wide range of biological specimens. Immobilization of biological samples was examined with a variety of adhesives. For histological sections, a filter system for flat embedding was developed. We validated ultra-thin embedding on laser marked slides for efficient, high-resolution CLEM. Target cells can be re-located within minutes in SEM without protracted searching and correlative investigations were reduced to a minimum of preparation steps, while still reaching highest resolution. The FIB/SEM milling procedure is facilitated and significantly accelerated as: (i) milling a ramp becomes needless, (ii) significant re-deposition of milled material does not occur; and (iii) charging effects are markedly reduced. By optimizing all technical parameters FIB/SEM stacks with 2 nm iso-voxels were achieved over thousands of sections, in a wide range of biological samples.
Characteristics of live parameters of the HS‐5 human bone marrow stromal cell line cocultured with the leukemia cells in hypoxia, for the studies of leukemia–stroma cross‐talk

The unique bone marrow microenvironment is created by stromal cells and such physical conditions as hypoxia. Both hypoxia and interactions with stromal cells have a significant impact on the biology of leukemia cells, changing their sensitivity to antileukemic therapies. Thus, it is crucial to introduce biological systems, which enable the investigation of leukemia–stroma cross‐talk and verification of novel therapies effectiveness under such bone marrow niche‐mimicking conditions. Here, we have established an experimental setup based on the hypoxic co‐culture of stromal cells with different cell lines derived from various leukemia patients. Flow cytometry enables simultaneous fluorescent tracking of viable cells and analysis of fundamental cellular processes, also to monitor the basal vital state of cells in the hypoxic co‐culture. This is critically important, as the stromal cells deliver a big variability of signals to protect leukemia cells and provide drug resistance. Therefore, keeping stromal cells at the healthy state is crucial during experimental procedures. In the proposed studies, viability, apoptosis, proliferation, ROS production, and mitochondrial membrane potential were monitored in both cell types, which were separated on the basis of the fluorescence of a cell tracker. We have shown that the proposed hypoxic co‐culture conditions do not affect basal live parameters of stromal cells, indicating the relevance of proposed model. Finally, we utilized this experimental setup to monitor the stroma‐mediated protection of leukemia cells from the imatinib‐induced cell death, which contributes to the leukemia progression and development of therapy resistance. Altogether, we recommend such flow cytometric strategy as an elementary screen of the vital state of stromal cells, which should be performed when using the co‐culture hypoxic models. The proposed approach can also be broadly used for other studies of the leukemia–stroma cross‐talk and of the part played by the leukemic microenvironment in drug screening studies.
Basolateral presence of the proinflammatory cytokine tumor necrosis factor ‐α and secretions from adipocytes and macrophages reduce intestinal sugar transport

We have previously demonstrated in Caco‐2 cells that tumor necrosis factor‐α (TNF‐α) inhibits sugar uptake, acting from the apical membrane, by decreasing the expression of the Na+‐glucose cotransporter SGLT1 in the brush border membrane. The goal was to investigate the hypothesis that TNF‐α from abdominal adipose tissue (adipocytes and macrophages) would decrease sugar and amino acid transport acting from the basolateral membrane of the enterocytes. TNF‐α placed in the basal compartment of Caco‐2 cells decreased α‐methyl‐ d‐glucose (αMG) and glutamine uptake. The apical medium derived from these Caco‐2 cells apically placed in another set of cells, also reduced sugar and glutamine transport. Reverse‐transcription polymerase chain reaction analysis demonstrated upregulation of TNF‐α, IL‐1β, and MCP1 expression in Caco‐2 cells exposed to basal TNF‐α. Similarly, αMG uptake was inhibited after Caco‐2 cells were incubated, in the basal compartment, with medium from visceral human mesenchymal stem cells‐derived adipocytes of overweight individuals. The apical medium collected from those Caco‐2 cells, and placed in the upper side of other set of cells, also decreased sugar uptake. Basal presence of medium derived from lipopolysaccharide‐activated macrophages and nonactivated macrophages decreased αMG uptake as well. Diet‐induced obese mice showed an increase in the visceral adipose tissue surrounding the intestine. In this physiological condition, there was a reduction on αMG uptake in jejunal everted rings.
Materielle Mitarbeiterbeteiligung: Geringe Verbreitung, aber hohe Intensität

Das Thema Mitarbeiterbeteiligung ist derzeit – wieder einmal – in aller Munde. Unlängst haben SPD und CDU/CSU ihre Konzepte zur Förderung der Mitarbei terbeteiligung vorgelegt (vgl. Kasten auf Seite 5). Gerade im Kontext die ser aktuellen Diskussion über mögliche staatliche Subventionen ist die Bedeutung der Mitarbeiterbeteiligung in der betrieblichen Praxis – auch in der zeitlichen Entwicklung – eine zentrale Größe. Die Ergebnisse einer umfassenden IABBetriebsbefragung liefern Informationen über die Verbreitung und die Intensität von Mitarbeiterbeteiligung in ostund westdeutschen Betrieben.
MiR-665 aggravates heart failure via suppressing CD34-mediated coronary microvessel angiogenesis

Background: Heart failure (HF) is a major public health problem worldwide. The development of HF was related to coronary microvessel dysfunction. Whether miRNAs participate in HF by regulating coronary microvessel function remain unclear. Methods: The potential targets of miR-665 were predicted by rnahybrid software, then verified through anti-Ago2 co-immunoprecipitation, Western blotting and luciferase reporter assays. rAAV9 system was used to manipulate the expression of miR-665 in vivo. Results: Significant increase of miR-665 was observed in endothelial cells of human heart with heart failure. In vitro over-expression of miR-665 in endothelial cells resulted in decreased proliferation but enhanced apoptosis. rAAV-mediated delivery of miR-665 reduced coronary microvessel angiogenesis and cardiac microvessel density, then further impaired cardiac function in vivo. Furthermore, CD34 was confirmed as one of the miR-665 targets. Consistently, re-expression of CD34 attenuated miR-665-mediated damage effects in vitro and in vivo. We also found that Sp1 regulated miR-665 expression in endothelial cells. Conclusion: Our findings demonstrated that miR-665 played an important role in heart failure via damaging coronary microvessel angiogenesis, and suggested that miRNA-based therapeutics may protect against coronary microvessel dysfunction and heart failure.
Skeletal Status, Body Composition, and Glycaemic Control in Adolescents with Type 1 Diabetes Mellitus

Background Disturbed bone turnover, osteoporosis, and increased fracture risk are late complications of insulin-dependent diabetes mellitus. Little is known about how far and to what extent can glycaemic control of type 1 diabetes mellitus (T1DM) prevent disturbances of bone health and body composition during the growth and maturation period. Objective The aim of this cross-sectional study was to compare the skeletal status outcomes and body composition between patients stratified by glycaemic control (1-year HbA1c levels) into well- and poorly-controlled subgroups in a population of T1DM adolescents, that is, <8% and ≥8%, respectively. Subjects and Methods Skeletal status and body composition were evaluated in 60 adolescents with T1DM (53.3% female; mean aged: 15.1 ± 1.9 years; disease duration: 5.1 ± 3.9 years) using dual energy X-ray absorptiometry (GE Prodigy). The results were compared to age- and sex-adjusted reference values for healthy controls. The calculated Z-scores of different metabolic control subgroups were compared. Clinical data was also assessed. Results As evidenced by Z-scores, patients with T1DM revealed a significantly lower TBBMD (total body bone mineral density), TBBMC (total body bone mineral content), S24BMD (bone mineral density of lumbar spine L2–L4), and TBBMC/LBM ratio (total body bone mineral content/lean body mass), but higher FM (fat mass) and FM/LBM ratio (fat mass/lean body mass) values compared to an age- and sex-adjusted general population. The subset (43.3% patients) with poor metabolic control (HbA1c ≥ 8%) had lower TBBMD, TBBMC, and LBM compared to respective values noted in the HbA1c < 8% group, after adjusting for confounders (mean Z-scores: −0.74 vs. −0.10, p = 0.037; −0.67 vs. +0.01, p = 0.026; and −0.45 vs. +0.20, p = 0.043, respectively). Additionally, we found a significant difference in the TBBMC/LBM ratio (relative bone strength index) between the metabolic groups (−0.58 vs. −0.07; p = 0.021). A statistically significant negative correlation between 1-year HbA1c levels and Z-scores of TBBMD, TBBMC, and LBM was also observed. In patients with longer disease duration, a significant negative correlation was established only for TBBMD, after adjusting for confounders. The relationships between densitometric values and age at onset of T1DM and sex were not significant and showed no relation to any of the analysed parameters of the disease course. Conclusion Findings from this study of adolescents with T1DM indicate that the lower Z-scores of TBBMD, TBBMC, and LBM as well as the TBBMC/LBM ratio are associated with increased HbA1c levels. Their recognition can be crucial in directing strategies to optimise metabolic control and improve diabetes management for bone development and maintenance in adolescents with T1DM.
Combined brain‐derived neurotrophic factor with extinction training alleviate impaired fear extinction in an animal model of post‐traumatic stress disorder

Impaired fear memory extinction (Ext) is one of the hallmark symptoms of post‐traumatic stress disorder (PTSD). However, since the precise mechanism of impaired Ext remains unknown, effective interventions have not yet been established. Recently, hippocampal‐prefrontal brain‐derived neurotrophic factor (BDNF) activity was shown to be crucial for Ext in naïve rats. We therefore examined whether decreased hippocampal‐prefrontal BDNF activity is also involved in the Ext of rats subjected to a single prolonged stress (SPS) as a model of PTSD. BDNF levels were measured by enzyme‐linked immunosorbent assay (ELISA), and phosphorylation of TrkB was measured by immunohistochemistry in the hippocampus and medial prefrontal cortex (mPFC) of SPS rats. We also examined whether BDNF infusion into the ventral mPFC or hippocampus alleviated the impaired Ext of SPS rats in the contextual fear conditioning paradigm. SPS significantly decreased the levels of BDNF in both the hippocampus and mPFC and TrkB phosphorylation in the ventral mPFC. Infusion of BDNF 24 hours after conditioning in the infralimbic cortex (ILC), but not the prelimbic cortex (PLC) nor hippocampus, alleviated the impairment of Ext. Since amelioration of impaired Ext by BDNF infusion did not occur without extinction training, it seems the two interventions must occur consecutively to alleviate impaired Ext. Additionally, BDNF infusion markedly increased TrkB phosphorylation in the ILC of SPS rats. These findings suggest that decreased BDNF signal transduction might be involved in the impaired Ext of SPS rats, and that activation of the BDNF‐TrkB signal might be a novel therapeutic strategy for the impaired Ext by stress.
Fear of Recurrence and View of Life Affect Health-Related Quality of Life in Patients with Differentiated Thyroid Carcinoma: A Prospective Swedish Population-Based Study.

BACKGROUND Differentiated thyroid cancer (DTC) has a good prognosis but a remaining risk of recurrence and need for lifelong follow-up. The aim was to study changes in health-related quality of life (HRQoL) from diagnosis to one year of follow-up. METHODS In this prospective population-based study, patients were identified from all the Swedish departments of oncology treating DTC. In total, 487 patients diagnosed with DTC between 2012 and 2017 were invited to answer the Swedish version of the Short Form-36 Health Survey (SF-36) and a study-specific questionnaire at diagnosis and after one year. RESULTS In total, 349 (72%) patients responded. Of these, 235 (67%) had reached one year of follow-up and also answered the questionnaire after one year. Of those, 23% had a negative view of life, and 75% had a fear of recurrence at diagnosis. These patients had a significantly lower HRQoL on eight and five of the SF-36 domains, respectively (p < 0.05). A negative view of life and a fear of recurrence also affected HRQoL negatively after one year of follow-up, with a significantly lower HRQoL on seven SF-36 domains for those reporting a negative view of life or fear of recurrence often (p < 0.05). Thyrotropin suppression did not affect HRQoL negatively. In regression models, HRQoL at diagnosis was the most important predictive factor for HRQoL at the one-year follow-up. CONCLUSIONS Despite a good prognosis, HRQoL was substantially affected at the time of diagnosis, with some improvements after one year. As fear of recurrence and a negative view of life substantially affect HRQoL, these patients should be given additional attention.
Evaluating an AAC training for special education teachers in Sri Lanka,a low- and middle-income country

Abstract A majority of people with disabilities live in low- and middle-income countries where communication services for individuals with complex communication needs are scarce. It is essential that communication partners of individuals who have complex communication needs receive training to support communication. To address this issue, the current study evaluated an augmentative and alternative communication training for special education teachers living in a low- and middle-income country, Sri Lanka. The training was based on four key practices identified when training in low- and middle-income countries: investigate learner needs, provide contextually relevant instructional content, use engaging instructional activities, and assess the impact of instructional activities. This investigation implemented an interrupted time-series design with nine teacher–student dyads. Special education teachers were taught to provide evocative communication opportunities. Following the training, all nine special education teachers significantly increased the number of evocative communication opportunities provided to students with complex communication needs. Additionally, all nine students with complex communication needs significantly increased their number of communication turns.
The Use of Regional or Local Anesthesia for Carotid Endarterectomies May Reduce Blood Loss and Pulmonary Complications.

OBJECTIVE Over 150,000 carotid endarterectomy (CEA) procedures are performed each year. Perioperative anesthetic management may be complex due to multiple patient and procedure-related risk factors. The authorsaimed to determine whether the use of general anesthesia (GA), when compared with regional anesthesia (RA), would be associated with reduced perioperative morbidity and mortality in patients undergoing a CEA. DESIGN Retrospective analysis of the American College of Surgeons National Surgical Quality Improvement Program (NSQIP) database. SETTING The authors evaluated patients undergoing a CEA at multiple university- and community-based settings. PARTICIPANTS A total of 43,463 patients were identified; 22,845 patients were propensity matched after excluding for missing data. INTERVENTIONS The study population was divided into 2 groups: patients undergoing RA or GA. The RA group included regional anesthesia performed by the anesthesiologist or surgeon, monitored anesthesia care, and local infiltration. METHODS The primary endpoint was 30-day mortality. Secondary endpoints included surgical site infection, pulmonary complications, return to the operating room, acute kidney injury, cardiac arrest, urinary tract infection, myocardial infarction, thromboembolism, perioperative transfusion, sepsis, and days to discharge. MEASUREMENTS AND MAIN RESULTS Younger age, Hispanic ethnicity, body mass index <18.5, dyspnea, chronic obstructive pulmonary disease, and smoking history were associated with receiving GA. Patients with low hematocrit and low platelets were more likely to get RA. There was no mortality difference. GA was associated with a significantly higher rate of perioperative transfusions (p = 0.037) and perioperative pneumonia (p = 0.027). CONCLUSION The use of RA over GA in CEA is associated with decreased risk of postoperative pneumonia and a reduced need for perioperative blood transfusions.
Induction of the pneumococcal vncRS operon by lactoferrin is essential for pneumonia

ABSTRACT Streptococcus pneumoniae (pneumococcus), the major pathogen for pneumonia, commonly colonizes the lung, but the mechanism underlying the coordination of virulence factors during invasion via the host protein remains poorly understood. Bacterial lysis releases the components of the cell wall, and triggers innate immunity and the subsequent secretion of pro-inflammatory cytokines. Previously, the virulence of the pep27 mutant was shown to be attenuated as a feasible candidate for vaccine development. However, the role of pep27 gene, belonging to the vancomycin-resistance locus (vncRS operon), in virulence, is largely unknown. This study demonstrates that transferrin in the host serum reduces the survival of the host during S. pneumoniae infections in mice. The exposure of the pneumococcal D39 strain to lactoferrin induced the vncRS operon, lysis, and subsequent in vivo cytokine production, resulting in lung inflammation. However, these responses were significantly attenuated in pneumococci harboring a mutation in pep27. Mechanistically, the VncS ligand, identified as lactoferrin, induced the vncRS operon and increased the in vivo mortality rates. Thus, serum-induced activation of vncRS plays an essential role in inducing pneumonia.
Adding Diversity and Culture to the Engineer’s Toolkit: Evaluating a Unique Course Option for Engineering Students

Lack of diversity in engineering is a persistent problem at both industry and collegiate levels. Defined for this paper as individual qualities divergent from historical characteristics (or even stereotypes) of engineers, diversity, at times, seems to be at odds with the culture of engineering itself. Lack of diversity in engineering manifests in a decreased retention rate of underrepresented minorities and women pursuing engineering degrees and engineering careers, a culture of homogeneity in engineering education and professional practice, and ensuing limitations to engineering design and product development. A more diverse engineering enterprise, by contrast, increases inclusivity and can provide a competitive advantage in industry [1]. One potential solution to this problem is the integration of diversity education into engineering curriculum at the undergraduate and graduate levels. Diversity education here includes courses addressing the culture and social context of engineering, the disparate outcomes of engineers belonging to groups traditionally underrepresented in engineering, and inclusive engineering practice. Courses such as these would help students contextualize their experiences and those of others in a larger body of knowledge about human interaction, challenge implicit biases, and make a statement about institutional values of inclusion. These courses may align with a larger movement in engineering education to integrate ethics, human-centered design, leadership development, and community-based project work—considerations of people, in other words— into more traditional technical coursework [2] [5]. This paper examines one such course offered at Stanford in 2017 as both a case study and reflection opportunity. This paper is authored by a student who was enrolled in the 2017 course, with the collaboration and co-authorship of the course instructors. Together our work relays the course experience from a first-hand perspective (through the experience of the lead author) and evaluates several course aspects, including its impact and efficacy (through instrumentation developed by the instructors and by the institution and administered to all students in the class). We share recommendations for course improvement, and for related curricular efforts in engineering, based on both student reflection and evaluation data. Our analysis and recommendations focus on inclusivity, which is the ability of students of all backgrounds to engage meaningfully with the course. Since the goal of such courses is to attract students with a range of experiences and backgrounds, inclusivity in every course component is critical.
Generation of a versatile BiFC ORFeome library for analyzing protein–protein interactions in live Drosophila

Transcription factors achieve specificity by establishing intricate interaction networks that will change depending on the cell context. Capturing these interactions in live condition is however a challenging issue that requires sensitive and non-invasive methods. We present a set of fly lines, called “multicolor BiFC library”, which covers most of the Drosophila transcription factors for performing Bimolecular Fluorescence Complementation (BiFC). The multicolor BiFC library can be used to probe binary or tripartite interactions and is compatible for large-scale interaction screens. The library can also be coupled with established Drosophila genetic resources to analyze interactions in the developmentally relevant expression domain of each protein partner. We provide proof of principle experiments of these various applications, using Hox proteins in the live Drosophila embryo as a case study. Overall this novel collection of ready-to-use fly lines constitutes an unprecedented genetic toolbox for the identification and analysis of protein-protein interactions in vivo.