Miracle-dz/UniFAR-FTUs huggingface dataset:
Organization and Myogenic Restricted Expression of the Murine Serum Response Factor Gene

Serum response factor (SRF), a member of an ancient family of DNA-binding proteins, is generally assumed to be a ubiquitous transcription factor involved in regulating growth factor-responsive genes. However, avian SRF was recently shown (Croissant, J. D., Kim, J.-H., Eichele, G., Goering, L., Lough, J., Prywes, R., and Schwartz, R. J. (1996) Dev. Biol. 177, 250–264) to be preferentially expressed in myogenic lineages and is required for regulating post-replicative muscle gene expression. Given the central importance of SRF for the muscle tissue-restricted expression of the striated α-actin gene family, we wanted to determine how SRF might contribute to this muscle-restricted expression. Here we have characterized the murine SRF genomic locus, which has seven exons interrupted by six introns, with the entire locus spanning 11 kilobases. Murine SRF transcripts were processed to two 3′-untranslated region polyadenylation signals, yielding 4.5- and 2.5-kilobase mRNA species. Murine SRF mRNA levels were the highest in adult skeletal and cardiac muscle, but barely detected in liver, lung, and spleen tissues. During early mouse development,in situ hybridization analysis revealed enrichment of SRF transcripts in the myotomal portion of somites, the myocardium of the heart, and the smooth muscle media of vessels of mouse embryos. Likewise, murine SRF promoter activity was tissue-restricted, being 80-fold greater in primary skeletal myoblasts than in liver-derived HepG2 cells. In addition, SRF promoter activity increased 6-fold when myoblasts withdrew from the cell cycle and fused into differentiated myotubes. A 310-base pair promoter fragment depended upon multiple intact serum response elements in combination with Sp1 sites for maximal myogenic restricted activity. Furthermore, cotransfected SRF expression vector stimulated SRF promoter transcription, whereas dominant-negative SRF mutants blocked SRF promoter activity, demonstrating a positive role for an SRF-dependent autoregulatory loop.
Effects of combining hygro-thermal stress on reliability of plastic QFN package

In this paper, finite element analysis (FEA) was used to evaluate the stress distribution on quad flat no-lead (QFN) packages. Different models were established, including moisture diffusion, thermo-mechanical stress, hygro-mechanical stress and hygro-thermo-mechanical stress models. The transient moisture absorption and desorption analysis were performed to estimate the package moisture distribution. A variety of experimental works were designed to study the hygro-thermal stress on the interface of die/mold compound and die attachment delaminaion. After the moisture preconditioning under 85°C/85%RH, it is found that the external mold compound (MC) material of package is almost fully saturated. The differential swelling that occurs between the mold compound and nonpolymeric materials leads to hygroscopic mismatch stresses in the package. The FEA results revealed that there is a significant contribution of hygroscopic swelling induced stress and an obvious effect of hygro-thermo-mechanical stress on package reliability. The experimental results also show that the hygro-thermo-mechanical stress under lead free reflow temperature produce considerable influence to the package integrity after moisture precondition.
Root extract of Prunella vulgaris inhibits in vitro and in vivo carcinogenesis in MCF-5 human breast carcinoma via suppression of angiogenesis, induction of apoptosis, cell cycle arrest and modulation of PI3K/AKT signalling pathway.

PURPOSE In this study we examined the anticancer effects of methanolic root extract of Prunella Vulgaris (PVE) against the MCF-5 breast cancer (BC) cell line along with its mode of action. METHODS The proliferation rate of the MCF-5 cells was assessed by MTT assay. Apoptosis was confirmed by acridine orange (AO)/ethidium bromide (EB) and annexin V/propidium iodide (PI) staining. DNA damage was checked by comet assay. Cell cycle analysis was performed by flow cytometry. Protein expression was determined by western blotting. In vivo evaluation of the extract was carried out in xenografted tumor mice models. RESULTS PVE inhibited the growth of the MCF-5 cells and exhibited an IC50 value of 25 µg/ml. The investigation of underlying mechanism revealed that PVE triggered apoptotic cell death of the MCF-5 cells which was also associated with enhancement of the expression of Bax and decrease in the expression of Bcl-2. PVE also caused arrest of the cells in the G2/M phase of the cell cycle and also exerted the anti-angiogenic effects. In vivo evaluation of PVE showed that it could inhibit the tumor weight and volume, suggestive of the anticancer potential of PVE. CONCLUSION The root extract of Prunella vulgaris in this study was shown to exert potent anticancer effects in MCF-7 human BC cells both in vitro and in vivo, accompanied with apoptosis induction, inhibition of angiogenesis, cell cycle arrest, and modulation of PI3K/AKT signaling pathway.
Role of GluN2A NMDA receptor in homocysteine‐induced prostaglandin E2 release from neurons

Hyperhomocysteinemia or systemic elevation of homocysteine is a metabolic condition that has been linked to multiple neurological disorders where inflammation plays an important role in the progression of the disease. However, it is unclear whether hyperhomocysteinemia contributes to disease pathology by inducing an inflammatory response. The current study investigates whether exposure of primary cultures from rat and mice cortical neurons to high levels of homocysteine induces the expression and release of the proinflammatory prostanoid, Prostaglandin E2 (PGE2). Using enzymatic assays and immunoblot analysis we show concurrent increase in the activity of cytosolic phospholipase A2 (cPLA2) and level of cyclooxygenase‐2 (COX2), two enzymes involved in PGE2 biosynthesis. The findings also show an increase in PGE2 release from neurons. Pharmacological inhibition of GluN2A‐containing NMDAR (GluN2A‐NMDAR) with NVP‐AAM077 significantly reduces homocysteine‐induced cPLA2 activity, COX2 expression, and subsequent PGE2 release. Whereas, inhibition of GluN2B‐containing NMDAR (GluN2A‐NMDAR) with Ro 25‐6981 has no effect. Complementary studies in neuron cultures obtained from wild type and GluN2A knockout mice show that genetic deletion of GluN2A subunit of NMDAR attenuates homocysteine‐induced neuronal increase in cPLA2 activity, COX2 expression, and PGE2 release. Pharmacological studies further establish the role of both extracellular‐regulated kinase/mitogen‐activated protein kinase and p38 MAPK in homocysteine‐GluN2A NMDAR‐dependent activation of cPLA2‐COX2‐PGE2 pathway. Collectively, these findings reveal a novel role of GluN2A‐NMDAR in facilitating homocysteine‐induced proinflammatory response in neurons.
Current treatment status and barriers for patients with chronic HCV infection in mainland China

Abstract Chronic hepatitis C virus (HCV) infection is a serious public health problem worldwide. China, as the country with the largest number of HCV infections in the world, plays a significant role in eliminating hepatitis C. Due to different financial situations and education background, hepatitis C patients take different actions for their disease treatment and management. Therefore, antiviral treatment status should be attached great importance to learn the medical demand of patients. A nationwide, multicenter survey was conducted from July 2015 to June 2016. Of 1798 inpatients and outpatients with chronic HCV from 56 hospitals participated in the survey. Each patient completed the questionnaire with questions about his/her antiviral therapy status, perception of treatment barriers, and expectations for future treatment. In total 1622 patients, including 1241 with chronic hepatitis C, 344 with cirrhosis, and 37 patients with hepatocellular carcinoma, fulfilled data collection requirements and finally were included in analysis. Overall, up to 30.7% of the patients had not or currently does not intend to receive antiviral therapy. The main reason was expecting more potent and well-tolerance medication (31.5%), followed by the fear of interferon related side effects (27.5%). Multiple regression analysis showed that the patient's annual income, the severity of HCV, and comorbidity were independent predictors of not receiving antiviral therapy. The whole patients were expecting more potent and well tolerance medication available soon. In summary, Peg-IFN/RBV treatment regimen cannot meet the need of patients well, and safe and efficient direct-acting antivirals are urgently needed in mainland China.
The risk of recurrent stroke at 24 months in patients with symptomatic carotid near‐occlusion: results from CAOS, a multicentre registry study

The risk of recurrent stroke amongst patients with symptomatic carotid near‐occlusion (SCNO) has not been clearly established, and its management remains controversial. The aim was to define the 24‐month risk of recurrent stroke and to analyse the effect of the different treatment modalities (medical treatment and revascularization) in a population of patients with SCNO.
The Effect of Fossil Sampling on the Estimation of Divergence Times With The Fossilised Birth Death Process.

Timescales are of fundamental importance to evolutionary biology as they facilitate hypothesis tests of historical evolutionary processes. Through the incorporation of fossil occurrence data, the fossilised birth-death (FBD) process provides a framework for estimating divergence times using more palaeontological data than traditional node calibration approaches have allowed. The inclusion of more data can refine evolutionary timescale estimates, but for many taxonomic groups it is computationally infeasible to include all fossil occurrence data. Here, we utilise both empirical data and a simulation framework to identify approaches to subsampling fossil occurrence data that result in the most accurate estimates of divergence times. To achieve this we assess the performance of the FBD-Skyline model when implementing multiple approaches to incorporating subsampled fossil occurrences. Our results demonstrate that it is necessary to account for all available fossil occurrence data to achieve the most accurate estimates of clade age. We show that this can be achieved if an empirical Bayes approach to account for fossil sampling through time is applied to the FBD process. Random subsampling of occurrence data can lead to estimates of clade age that are incompatible with fossil evidence if no control over the affinities of fossil occurrences is enforced. Our results call into question the accuracy of previous divergence time studies incorporating the FBD process that have used only a subsample of all available fossil occurrence data.
Antigenic Properties and Diagnostic Potential of Baculovirus-Expressed Infectious Bursal Disease Virus Proteins VPX and VP3

ABSTRACT The routine technique for detecting antibodies specific to infectious bursal disease virus (IBDV) is a serological evaluation by enzyme-linked immunosorbent assay (ELISA) with preparations of whole virions as the antigens. To avoid using complete virus in the standard technique, we have developed two new antigens through the expression of the VPX and VP3 genes in insect cells. VPX and especially VP3 were expressed at high levels in insect cells and simple to purify. The immunogenicity of both proteins was similar to that of the native virus. VPX was able to elicit neutralizing antibodies but VP3 was not. Purified VPX and VP3 were tested in an indirect ELISA with more than 300 chicken sera. There was an excellent correlation between the results of the ELISA using VPX and those of the two commercial kits. VP3 did not perform as well as VPX, and the linear correlation was significantly lower. A comparison with the standard reference technique, seroneutralization, showed that the indirect ELISA was more sensitive. Therefore, VPX-based ELISA is a good alternative to conventional ELISAs that use whole virions.
Modulation of macrophage subtypes by IRF5 determines osteoclastogenic potential

Bone‐resorbing osteoclasts are differentiated from macrophages (MΦ) by M‐CSF and RANKL. MΦ can be mainly classified into M1 and M2 MΦ, which are proinflammatory and anti‐inflammatory, respectively, but little is known about their osteoclastogenic potential. Here, we investigated the osteoclastogenic potential of MΦ subtypes. When the two MΦ subtypes were differentiated into osteoclasts using M‐CSF and RANKL, M2 MΦ more potently differentiated into osteoclasts than M1 MΦ. M2 MΦ generated with IL‐4 or IL‐10 also showed enhanced osteoclast differentiation compared with M1 MΦ induced by IFN‐γ and lipopolysaccharide. In addition, robust bone‐resorptive capacity and giant actin rings, which are features of mature osteoclasts, were observed in M2, but not M1 MΦ, under the osteoclast differentiation condition. Osteoclast differentiation was significantly increased in CD206+ M2 MΦ but not in CD86+ M1 MΦ. Compared with M1 MΦ, c‐Fms and RANK were highly expressed in M2 MΦ. Enhanced osteoclastogenesis of M2 MΦ was mediated through sustained ERK activation, followed by efficient c‐Fos and NFATc1 induction. Notably, the osteoclastogenic potential of M1 MΦ converted into M2 MΦ by exposure to M‐CSF was higher than that of M2 MΦ converted into M1 MΦ by exposure to GM‐CSF. Silencing IRF5, which is responsible for M1 MΦ polarization, increased osteoclast differentiation by enhancing c‐Fms expression and activation of ERK, c‐Fos, CREB, and NFATc1, which was inhibited by overexpression of IRF5. Collectively, M2 MΦ are suggested to be more efficient osteoclast precursors than M1 MΦ because of the attenuated expression of IRF5.
Regulation of Diabetogenic Immunity by IL-15–Activated Regulatory CD8 T Cells in Type 1 Diabetes

Key Points NOD mice are deficient in number and function of CD8 Tregs that control the germinal center. Treatment with IL-15 superagonist expands NOD CD8 Tregs and restores their activity. IL-15–activated CD8 Tregs delay the adoptive transfer of type 1 diabetes. Unchecked collaboration between islet-reactive T and B lymphocytes drives type 1 diabetes (T1D). In the healthy setting, CD8 T regulatory cells (Tregs) terminate ongoing T–B interactions. We determined that specific CD8 Tregs from NOD mice lack suppressive function, representing a previously unreported regulatory cell deficit in this T1D-prone strain. NOD mice possess 11-fold fewer Ly-49+ CD8 Tregs than nonautoimmune mice, a deficiency that worsens as NOD mice age toward diabetes and leaves them unable to regulate CD4 T follicular helper cells. As IL-15 is required for Ly-49+ CD8 Treg development, we determined that NOD macrophages inadequately trans-present IL-15. Despite reduced IL-15 trans-presentation, NOD Ly-49+ CD8 Tregs can effectively transduce IL-15–mediated survival signals when they are provided. Following stimulation with an IL-15/IL-15Ra superagonist complex, Ly-49+ CD8 Tregs expanded robustly and became activated to suppress the Ag-specific Ab response. IL-15/IL-15Ra superagonist complex–activated CD8+CD122+ T cells also delayed diabetes transfer, indicating the presence of an underactivated CD8 T cell subset with regulatory capacity against late stage T1D. We identify a new cellular contribution to anti-islet autoimmunity and demonstrate the correction of this regulatory cell deficit. Infusion of IL-15–activated CD8 Tregs may serve as an innovative cellular therapy for the treatment of T1D.
OATP1B3-1B7 (LST-3TM12) Is a Drug Transporter That Affects Endoplasmic Reticulum Access and the Metabolism of Ezetimibe

Drug transporters play a crucial role in pharmacokinetics. One subfamily of transporters with proven clinical relevance are the OATP1B transporters. Recently we identified a new member of the OATP1B family named OATP1B3-1B7 (LST-3TM12). This functional transporter is encoded by SLCO1B3 and SLCO1B7. OATP1B3-1B7 is expressed in hepatocytes and is located in the membrane of the smooth endoplasmic reticulum (SER). One aim of this study was to test whether OATP1B3-1B7 interacts with commercial drugs. First, we screened a selection of OATP1B substrates for inhibition of OATP1B3-1B7–mediated transport of dehydroepiandrosterone sulfate and identified several inhibitors. One such inhibitor was ezetimibe, which not only inhibited OATP1B3-1B7 but is also a substrate, as its cellular content was significantly increased in cells heterologously expressing the transporter. In humans, ezetimibe is extensively metabolized by hepatic and intestinal uridine-5′-diphospho-glucuronosyltransferases (UGTs), the catalytic site of which is located within the SER lumen. After verification of OATP1B3-1B7 expression in the small intestine, we determined in microsomes whether SER access can be modulated by inhibitors of OATP1B3-1B7. We were able to show that these compounds significantly reduced accumulation in small intestinal and hepatic microsomes, which influenced the rate of ezetimibe β-D-glucuronide formation as determined in microsomes treated with bromsulphthalein. Notably, this molecule not only inhibits the herein reported transporter but also other transport systems. In conclusion, we report that multiple drugs interact with OATP1B3-1B7; for ezetimibe, we were able to show that SER access and metabolism is significantly reduced by bromsulphthalein, which is an inhibitor of OATP1B3-1B7. Significance Statement OATP1B3-1B3 (LST-3TM12) is a transporter that has yet to be fully characterized. We provide valuable insight into the interaction potential of this transporter with several marketed drugs. Ezetimibe, which interacted with OATP1B3-1B7, is highly metabolized by uridine-5′-diphospho-glucuronosyltransferases (UGTs), whose catalytic site is located within the smooth endoplasmic reticulum (SER) lumen. Through microsomal assays with ezetimibe and the transport inhibitor bromsulphthalein we investigated the interdependence of SER access and the glucuronidation rate of ezetimibe. These findings led us to the hypothesis that access or exit of drugs to the SER is orchestrated by SER transporters such as OATP1B3-1B7.
Development of a 3 T 3-like Line from an Embryo Culture of an Inbred Strain of Syrian Golden Hamster 1

Addition of fresh medium containing 5 or 10% fetal calf serum to a confluent culture induced DNA synthesis in 18 hr with subsequent cell division. Cells were hyperdiploid with a mode of 45 chromosomes (80% of the cells). When cells at the 60th passage were injected into the skin or cheek pouch of an inbred hamster of the same strain as that from which they were derived, they produced a benign tumor that regressed after 3 weeks. Morphological transformation was obtained by infection with the Moloney strain of murine sarcoma virus.
Differential cell analysis of cytocentrifuged bronchoalveolar fluid samples affected by the area counted.

OBJECTIVE To investigate variations in the differential cell counts between the quadrants of cytocentrifuged bronchoalveolar lavage (BAL) fluid preparations and to evaluate the diagnostic impact of these differences in interstitial lung diseases (ILD). STUDY DESIGN BAL fluid samples obtained from 30 patients suspected of having ILD or pneumonia were cytocentrifuged and additionally stained with May-Grünwald-Giemsa stain. Two observers differentiated 200 cells in each quadrant as well as in a circular pattern around the center of the cytocentrifuge spot. RESULTS Lymphocytes and alveolar macrophages were not randomly distributed on the cytocentrifuge spot. Ten samples of patients with histologically confirmed ILD were selected to test the diagnostic impact using a validated computer program. The predicted diagnosis did not correspond to the histologic diagnosis for one quadrant from 1 of these 10 samples (sarcoidosis instead of idiopathic pulmonary fibrosis), whereas the differential cell counts performed around the center of the cytocentrifuge spot provided the correct diagnosis in all cases. CONCLUSION BAL fluid differential cell counts varied between the quadrants of the cytocentrifuge spot. The center of the cytocentrifuge spot appeared to be the most reliable area. Therefore, cell counting is recommended in a circular pattern around the center of the cytocentrifuge spot.
Acute myocardial infarction: Does survival depend on geographical location and social background?

Aims This study described the interplay between geographical and social inequalities in survival after incident acute myocardial infarction (AMI) and examined whether geographical variation in survival exists when accounting for sociodemographic characteristics of the patients and their neighbourhood. Methods Ringmap visualization and generalized linear models were performed to study post-AMI mortality. Three individual-level analyses were conducted: immediate case fatality, mortality between days 1 and 28 after admission and 365-day survival among patients who survived 28 days after admission. Results In total, 99,013 incident AMI cases were registered between 2005 and 2014 in Denmark. Survival after AMI tended to correlate with sociodemographic indicators at the municipality level. In individual-level models, geographical inequality in immediate case fatality was observed with high mortality in northern parts of Jutland after accounting for sociodemographic characteristics. In contrast, no geographical variation in survival was observed among patients who survived 28 days. In all three analyses, odds and rates of mortality were higher among patients with low educational level (odds ratio (OR) (95% credible intervals) of 1.20 (1.12–1.29), OR of 1.12 (1.01–1.24) and mortality rate ratio of 1.45 (1.30–1.61)) and low income (OR of 1.24 (1.15–1.33), OR of 1.33 (1.20–1.48) and mortality rate ratio of 1.25 (1.13–1.38)). Conclusion Marked geographical inequality was observed in immediate case fatality. However, no geographically unequal distribution of survival was found among patients who survived 28 days after AMI. Results additionally showed social inequality in survival following AMI.
Independent ischemic stroke risk factors in older Americans: a systematic review

The Framingham Stroke Risk Profile (FSRP) is a validated model for predicting 10-year ischemic stroke risk in middle-aged adults, yet has not been demonstrated to consistently translate in older populations. This is a systematic review of independent risk factors measured among > 65 year olds, with subsequent first ischemic stroke, using PRISMA guidelines. We appraised peer-reviewed publications that included participants > 65 years old at risk assessment. Combined with other criteria, results were abstracted from 28 papers reporting six types of stroke risk factors: Serologic/Diagnostic, Conventional, Psychosocial, Genetic, Cognitive, and Antibiotic use. These studies demonstrated levels of serum androgens, C-reactive protein, and advanced glycation endproducts; thrombin generation; left ventricular mass; depressive symptoms; phosphodiesterase 4D single nucleotide polymorphisms; coagulation factor XII gene; peak thrombus generation; and lower cognitive functioning were independent risk factors for ischemic stroke in older adults. Plasma adipokines, free fatty acids and antibiotic use did not predict ischemic stroke. Purpose in life and APOEε2 allele were protective for ischemic stroke. This systematic review provides evidence of risk and protective factors for ischemic stroke in older cohorts that are not included in the FSRP. Further studies are needed to understand whether these factors are important enough to comprise a risk score.