Miracle-dz/UniFAR-FTUs huggingface dataset:
Revealing cellular and molecular transitions in neonatal germ cell differentiation using single cell RNA sequencing

ABSTRACT Neonatal germ cell development provides the foundation of spermatogenesis. However, a systematic understanding of this process is still limited. To resolve cellular and molecular heterogeneity in this process, we profiled single cell transcriptomes of undifferentiated germ cells from neonatal mouse testes and employed unbiased clustering and pseudotime ordering analysis to assign cells to distinct cell states in the developmental continuum. We defined the unique transcriptional programs underlying migratory capacity, resting cellular states and apoptosis regulation in transitional gonocytes. We also identified a subpopulation of primitive spermatogonia marked by CD87 (plasminogen activator, urokinase receptor), which exhibited a higher level of self-renewal gene expression and migration potential. We further revealed a differentiation-primed state within the undifferentiated compartment, in which elevated Oct4 expression correlates with lower expression of self-renewal pathway factors, higher Rarg expression, and enhanced retinoic acid responsiveness. Lastly, a knockdown experiment revealed the role of Oct4 in the regulation of gene expression related to the MAPK pathway and cell adhesion, which may contribute to stem cell differentiation. Our study thus provides novel insights into cellular and molecular regulation during early germ cell development. Summary: Single cell transcriptomic analysis of undifferentiated germ cells in neonatal mouse testes reveals heterogeneity of gene expression across subpopulations and time, identifying potential novel transcriptional regulators during germ cell differentiation.
Methylation of progesterone receptor isoform A promoter in normal breast tissue: An epigenetic link between early age at menarche and risk of breast cancer?

Emerging evidence indicates that some altered patterns of methylation that occur in breast tumors may also be found in breast tissue of healthy women in relation to the breast cancer (BC) risk factors. Progesterone receptor (PR) isoform α is a crucial regulator of breast hormone responsiveness and its hypermethylation plays an important role in the initiation and development of breast tumors. However, such a methylation change in healthy women and its link with the different risk factors has not yet been investigated. In the present study, we aimed to examine the relationship of possible methylation changes within a critical region in the promoter CpG island of PGR‐α (progesterone receptor α) gene in the healthy women with a set of reproductive and nonreproductive BC risk factors. The breast tissues were collected from 120 cancer‐free women who had undergone cosmetic mammoplasty. The genomic DNA was extracted from the breast tissues and the methylation level of PGR‐α promoter CpG island was determined by using MeDIP‐qPCR assay. Using regression analysis, we found that increasing menarche age is inversely associated with the high methylation of PGR‐α promoter ( β = −0.790, SE = 0.362; P = 0.031). Although lactating women had more methylation than nonlactating women (P = 0.026, the t test), this result was not confirmed by regression models. Such an observation may be helpful in better understanding of the underlying mechanisms by which early age at menarche increases the risk of BC. However, this perspective requires further validations in larger studies of more subjects as well as the inclusion of other related genes.
The Link Between School Attendance and Good Health

More than 6.5 million children in the United States, approximately 13% of all students, miss 15 or more days of school each year. The rates of chronic absenteeism vary between states, communities, and schools, with significant disparities based on income, race, and ethnicity. Chronic school absenteeism, starting as early as preschool and kindergarten, puts students at risk for poor school performance and school dropout, which in turn, put them at risk for unhealthy behaviors as adolescents and young adults as well as poor long-term health outcomes. Pediatricians and their colleagues caring for children in the medical setting have opportunities at the individual patient and/or family, practice, and population levels to promote school attendance and reduce chronic absenteeism and resulting health disparities. Although this policy statement is primarily focused on absenteeism related to students’ physical and mental health, pediatricians may play a role in addressing absenteeism attributable to a wide range of factors through individual interactions with patients and their parents and through community-, state-, and federal-level advocacy.
Quantitative in vitro comparison of the thrombogenicity of commercial dental implants.

BACKGROUND Dental implants often have surface modifications that alter surface topography and chemistry to improve osseointegration and thereby increase treatment predictability. Surface contact-induced blood coagulation is associated with the onset of osseointegration. PURPOSE To quantitatively evaluate the thrombogenicity of two commercially available dental implants that have similar surface roughness but different surface chemistry. MATERIAL AND METHODS Two commercially available dental implants with anodized or sandblasted acid-etched surfaces were evaluated for thrombogenic properties. Thrombogenicity was assessed by incubating implants for 1 hour in fresh, partially heparinized blood followed by hemocyte quantification, microscopic evaluation, and quantification of thrombogenic biomarkers. RESULTS Fibrin coverage was significantly higher on the anodized surface compared with the sandblasted acid-etched surface (P < 0.0001). Platelet and white blood cell attachment followed a similar pattern. The increased thrombogenicity was confirmed based on a significant increase in the levels of the coagulation cascade biomarkers, thrombin antithrombin complex, and β-thromboglobulin (all P < 0.05). CONCLUSION Dental implants with comparable roughness but differing surface chemistry had differing extents of blood contact activation. These data suggest that surface chemistry from anodization augments implant thrombogenicity compared with that from sandblasting and acid-etching, which could have implications for osseointegration.
Licochalcone A Inhibits Cellular Motility by Suppressing E-cadherin and MAPK Signaling in Breast Cancer

A compound isolated from Glycyrrhiza uralensis, licochalcone A (LA) exhibits anti-inflammatory and anti-tumor properties in various cell lines. LA has been found to promote autophagy and suppress specificity protein 1, inducing apoptosis in breast cancer cells. However, the regulation of breast cancer cell invasion and migration by LA is elusive. Thus, the present study investigated whether LA induces apoptosis and cellular motility in MDA-MB-231 breast cells, and investigated the underlying molecular mechanisms. MDA-MB-231 cells treated with LA and cell viability measured by cell counting kit-8 assay. Apoptotic signal proteins checked by flow cytometry, fluorescent staining, and Western blot. LA effectively suppressed cell migration, and modulated E-cadherin and vimentin expression by blocking MAPK and AKT signaling. LA inhibited cell proliferation and cell cycle, modulated mitochondrial membrane potential and DNA damage, and reduced oxidative stress in MDA-MB-231 cells. LA also activated cleaved-caspase 3 and 9, significantly decreased Bcl-2 expression, ultimately causing the release of cytochrome c from the mitochondria into the cytoplasm. Overall, our findings suggest that LA decreases cell proliferation and increases reactive oxygen species production for induced apoptosis, and regulates E-cadherin and vimentin by reducing MAPK and AKT signaling, resulting in suppressed MDA-MB-231 cell migration and invasion.
Influence of the patellar button thickness on the knee flexion after total knee arthroplasty.

PURPOSE One of the main problems of knee replacement is the limit of knee flexion. This study focuses on the knee implant and the patellar component currently in use in total knee arthroplasty, analyzing the influence of patellar thickness on the degree of knee flexion following surgery. METHODS A kinematics study was performed to evaluate whether an optimal patellar thickness can be identified, which enables the maximum flexion angle to be achieved. Using TC images, a healthy model was built. On this basis, a model of a knee joint which had undergone total knee arthroplasty using a Legion PS prosthesis was constructed. Initially, the standard thickness of patellar implant (9 mm) was used to build the model; then several different patellar implant thicknesses (in the range of 5-15 mm) were analyzed. RESULTS The results show a non-linear trend: a button thickness of less than 9 mm does not change the flexion angle, whereas a button thickness of over 9 mm results in a loss of flexion. The flexion loss is significant in the first two additions of thicknesses but negligible in the last ones. CONCLUSIONS In the case studied, flexion reduction is not linearly proportional to the patellar thickness. The outcome of total knee arthroplasty is considered to be satisfactory with the standard patellar button. The results of this study could be used to compare the kinematics with other total prosthesis and patellar implants, and should enable the optimization of the patellar residue bone thickness to obtain deep flexion.
Cholecystokinin release triggered by NMDA receptors produces LTP and sound–sound associative memory

Significance Neuropeptides represent the most diverse family of neurotransmitters with a wide distribution in the rodent brain. Cholecystokinin (CCK) is an octapeptide present in very high concentrations in the cortex, where it is expressed in several circuits. It has emerged as a useful marker, for example as an interneuron subpopulation, but its function is still unclear. Here we show that CCK, via its CCKB receptor, is essential for high-frequency stimulation-induced long-term potentiation, and that release of CCK is controlled by NMDA receptors. Thus, CCK may play an important role in memory formation. Memory is stored in neural networks via changes in synaptic strength mediated in part by NMDA receptor (NMDAR)-dependent long-term potentiation (LTP). Here we show that a cholecystokinin (CCK)-B receptor (CCKBR) antagonist blocks high-frequency stimulation-induced neocortical LTP, whereas local infusion of CCK induces LTP. CCK−/− mice lacked neocortical LTP and showed deficits in a cue–cue associative learning paradigm; and administration of CCK rescued associative learning deficits. High-frequency stimulation-induced neocortical LTP was completely blocked by either the NMDAR antagonist or the CCKBR antagonist, while application of either NMDA or CCK induced LTP after low-frequency stimulation. In the presence of CCK, LTP was still induced even after blockade of NMDARs. Local application of NMDA induced the release of CCK in the neocortex. These findings suggest that NMDARs control the release of CCK, which enables neocortical LTP and the formation of cue–cue associative memory.
TMEM16A as a Potential Biomarker in the Diagnosis and Prognosis of Lung Cancer.

BACKGROUND Transmembrane protein 16A (TMEM16A), also known as ANO1 (anoctamin-1), was reported to be vital in the growth and invasion of several malignancies. However, role of TMEM16A in lung cancer remained unclear. The aim of this study was to evaluate the expression of TMEM16A and its significance in lung cancer. METHODS qRT-PCR and Western blots were performed to evaluate the TMEM16A mRNA and protein expression. Proliferation and invasion of H1299 cancer cells were evaluated by CCK-8 and transwell assays. Tumor volumes in nude mice implanted with H1299 cells were assessed once every week for 5 weeks by measuring 2 perpendicular dimensions. Immunofluorescent staining revealed expression of TMEM16A in nude mice cancer tissues. RESULTS Our findings provided compelling evidence that TMEM16A production in H1299 cells is 2.1 times higher than observations in HBE16 cells. We showed that overexpression of TMEM16A contributed to the proliferation of H1299 cells. Moreover, T16Ainh-A01, a specific TMEM16A inhibitor or shRNA targeting TMEM16A somewhat inhibited lung tumor cell growth and invasion as evident from in vitro studies and from in vivo xenograft-tumor growth. Inhibition of TMEM16A strongly suppressed EGFR phosphorylation and growth of lung cancer cells. Furthermore, a reduction of p-RAS and p-ERK1/2 was also observed. CONCLUSION TMEM16A promoted growth and invasion in lung cancer cells via an EGFR/ MAPK-dependent signaling pathway. So we infer TMEM16A membrane protein may have potential to serve as a biomarker in lung cancer.
Circulating tumor DNA dynamics using patient-customized assays are associated with outcome in neoadjuvantly treated breast cancer

Pathological complete response (pCR) is an accurate predictor of good outcome following neoadjuvant chemotherapy (NAC) for locally advanced breast cancer. The presence of circulating-tumor DNA (ctDNA) has recently been reported to be strongly predictive of poor outcome in similar patient groups. We monitored ctDNA levels from 10 women undergoing NAC for locally advanced breast cancer using a patient-specific, hybrid-capture sequencing technique sensitive to the level of one altered allele in 10,000. Plasma was collected prior to the start of NAC, prior to each infusion of NAC, and during follow-up for between 350 and 1150 d after the start of NAC. Prior to the start of NAC, ctDNA was detectable in 3/3 triple negative, 3/3 HER2+, and 2/4 HER2−, ER+ breast cancer patients. Total cell-free DNA levels were considerably higher when patients were on NAC than at other times. ctDNA dynamics during NAC showed that patients with pCR experienced rapid declines in ctDNA levels, whereas patients without pCR typically showed evidence of residual ctDNA after initiation of treatment. Intriguingly, two of three patients that showed marked increases in ctDNA while on NAC experienced rapid recurrences (<2 yr following start of NAC). The third patient that had increases in ctDNA levels while on NAC had low-grade ER+ disease and showed residual ctDNA after surgery, which became undetectable after local radiation. Taken together, these results demonstrate the ability of our approach to sensitively serially monitor ctDNA during NAC, and identifies a need to further investigate the possibility of stratifying patients who need additional treatment or identify therapies that are ineffective.
Similar Weight Loss Induces Greater Improvements in Insulin Sensitivity and Liver Function among Individuals with NAFLD Compared to Individuals without NAFLD

Background: Preliminary evidence suggests that weight loss among obese has differential metabolic effects depending on the presence of non-alcoholic fatty liver disease (NAFLD). We assessed whether NAFLD predisposes to differential changes in liver fat content, liver function, and metabolic parameters upon diet-induced weight loss in a 50-week intervention trial. Methods: 143 overweight and obese non-smokers underwent a 12-week dietary intervention and a 38-week follow-up. Diet-induced changes in anthropometric measures, circulating biomarkers, and magnetic resonance (MR)-derived liver fat content and adipose tissue volumes were evaluated by mixed linear models stratifying by NAFLD at baseline. Results: The prevalence of NAFLD at baseline was 52%. Diet-induced weight loss after 12 (NAFLD: 4.8 ± 0.5%, No NAFLD: 5.1 ± 0.5%) and 50 weeks (NAFLD: 3.5 ± 0.7%, No NAFLD: 3.5 ± 0.9%) was similar in both groups, while the decrease in liver fat was significantly greater in the NAFLD group (week 12: 32.9 ± 9.5% vs. 6.3 ± 4.0%; week 50: 23.3 ± 4.4% vs. 5.0 ± 4.2%). Decreases in biomarkers of liver dysfunction (GGT, ALT, AST) and HOMA IR were also significantly greater in the NAFLD group. Other metabolic parameters showed no significant differences. Conclusion: Our data suggest that individuals with NAFLD show greater improvements of liver function and insulin sensitivity after moderate diet-induced weight loss than individuals without NAFLD.
Empathy, Embodied Simulation, and the Brain: Commentary on Aragno and Zepf/Hartmann

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Cetrorelix, a Gonadotropin-Releasing Hormone Antagonist, Induces the Expression of Melatonin Receptor 1a in the Gonadotropin-Releasing Hormone Neuronal Cell Line GT1–7

Melatonin has been implicated in the control of the reproductive system, and the modulatory actions of melatonin on gonadotropin-releasing hormone (GnRH) neurons have been assumed to be indirectly mediated through afferent neurons. However, our previous studies demonstrate sexually dimorphic modulation of A-type γ-aminobutyric acid (GABA) receptor (GABAAR) currents by melatonin in adult rat GnRH neurons and a preferential expression of melatonin 1a receptor (MT1) in male GnRH neurons. Using immortalized GnRH neurons (GT1–7 cells), the present study investigated the mechanism by which the expression of melatonin receptors is regulated in GnRH neurons. Like endogenous GnRH neurons, GT1–7 cells express both GnRH and GnRH receptor mRNAs, indicating that the cells have a self-stimulatory system. A 2-iodomelatonin binding assay and RT-PCR analysis demonstrated that the cells expressed neither MT1 nor MT2. However, treatment of GT1–7 cells with the GnRH antagonist cetrorelix significantly increased 2-iodomelatonin binding and induced a time- and concentration-dependent MT1 mRNA expression. The GABAAR currents were then measured using a perforated patch-clamp technique to examine whether the treatment with cetrorelix changed the responses to melatonin. Melatonin augmented the GABAAR currents in GT1–7 cells treated with 1 μM cetrorelix for 24 h, while melatonin decreased the currents in the cells not treated with cetrorelix, probably via receptor-independent processes. The present results suggest that GnRH downregulates the expression of MT1 via an autocrine-paracrine mechanism in GT1–7 cells, and modifies the melatonin-induced modulation of GABAAR currents. These findings may provide one possible mechanism for the sexually dimorphic responses to melatonin in adult rat GnRH neurons.
Particulate Mass and Nonvolatile Particle Number Emissions from Marine Engines Using Low-Sulfur Fuels, Natural Gas, or Scrubbers

In order to meet stringent fuel sulfur limits, ships are increasingly utilizing new fuels or, alternatively, scrubbers to reduce sulfur emissions from the combustion of sulfur-rich heavy fuel oil. The effects of these methods on particle emissions are important, because particle emissions from shipping traffic are known to have both climatic and health effects. In this study, the effects of lower sulfur level liquid fuels, natural gas (NG), and exhaust scrubbers on particulate mass (PM) and nonvolatile particle number (PN greater than 23 nm) emissions were studied by measurements in laboratory tests and in use. The fuel change to lower sulfur level fuels or to NG and the use of scrubbers significantly decreased the PM emissions. However, this was not directly linked with nonvolatile PN emission reduction, which should be taken into consideration when discussing the health effects of emitted particles. The lowest PM and PN emissions were measured when utilizing NG as fuel, indicating that the use of NG could be one way to comply with up-coming regulations for inland waterway vessels. Low PN levels were associated with low elemental carbon. However, a simultaneously observed methane slip should be taken into consideration when evaluating the climatic impacts of NG-fueled engines.
The Association of Telomere Length in Peripheral Blood Cells with Cancer Risk: A Systematic Review and Meta-analysis of Prospective Studies

The association between telomere length (TL) in peripheral blood cells and cancer risk remains inconclusive. We carried out a meta-analysis on prospective studies. The study-specific RR estimates were first transformed to a common comparable scale and then were pooled by a random-effects model. The dataset was composed of 13,894 cases and 71,672 controls from 28 studies in 25 articles. In the comparison of the longest versus shortest third of TL, we observed a marginally positive association between longer TL and higher risk of total cancers [OR = 1.086; 95% confidence interval (CI), 0.952–1.238]. Subgroup analyses showed that the association was stronger in lung cancer (n = 3; OR = 1.690; 95% CI, 1.253–2.280), in men (n = 6; OR = 1.302; 95% CI, 1.120–1.514) and in studies with more precise methods for DNA extraction (phenol–chloroform, salting-out or magnetic bead, n = 6, OR = 1.618; 95% CI, 1.320–1.985) and TL measurement (multiplex Q-PCR, n = 8; OR = 1.439; 95% CI, 1.118–1.852). Our meta-analysis suggested longer TL in peripheral blood cells is a likely risk factor for lung cancer or cancers in men. Accurate DNA extraction and TL measurement methods make it more liable to find significant associations between TL and cancer risk and thus should be taken into consideration in future epidemiologic studies. Cancer Epidemiol Biomarkers Prev; 26(9); 1381–90. ©2017 AACR.
Stimulation of sphingosine 1-phosphate signaling as an alveolar cell survival strategy in emphysema.

RATIONALE Vascular endothelial growth factor receptor (VEGFR) inhibition increases ceramides in lung structural cells of the alveolus, initiating apoptosis and alveolar destruction morphologically resembling emphysema. The effects of increased endogenous ceramides could be offset by sphingosine 1-phosphate (S1P), a prosurvival by-product of ceramide metabolism. OBJECTIVES The aims of our work were to investigate the sphingosine-S1P-S1P receptor axis in the VEGFR inhibition model of emphysema and to determine whether stimulation of S1P signaling is sufficient to functionally antagonize alveolar space enlargement. METHODS Concurrent to VEGFR blockade in mice, S1P signaling augmentation was achieved via treatment with the S1P precursor sphingosine, S1P agonist FTY720, or S1P receptor-1 (S1PR1) agonist SEW2871. Outcomes included sphingosine kinase-1 RNA expression and activity, sphingolipid measurements by combined liquid chromatography-tandem mass spectrometry, immunoblotting for prosurvival signaling pathways, caspase-3 activity and terminal deoxynucleotidyltransferase-mediated dUTP nick end labeling assays, and airspace morphometry. MEASUREMENTS AND MAIN RESULTS Consistent with previously reported de novo activation of ceramide synthesis, VEGFR inhibition triggered increases in lung ceramides, dihydroceramides, and dihydrosphingosine, but did not alter sphingosine kinase activity or S1P levels. Administration of sphingosine decreased the ceramide-to-S1P ratio in the lung and inhibited alveolar space enlargement, along with activation of prosurvival signaling pathways and decreased lung parenchyma cell apoptosis. Sphingosine significantly opposed ceramide-induced apoptosis in cultured lung endothelial cells, but not epithelial cells. FTY720 or SEW2871 recapitulated the protective effects of sphingosine on airspace enlargement concomitant with attenuation of VEGFR inhibitor-induced lung apoptosis. CONCLUSIONS Strategies aimed at augmenting the S1P-S1PR1 signaling may be effective in ameliorating the apoptotic mechanisms of emphysema development.