Miracle-dz/UniFAR-FTUs huggingface dataset:
Familial Hypertrophic Cardiomyopathy Mutations in the Regulatory Light Chains of Myosin Affect Their Structure, Ca2+Binding, and Phosphorylation*

The effect of the familial hypertrophic cardiomyopathy mutations, A13T, F18L, E22K, R58Q, and P95A, found in the regulatory light chains of human cardiac myosin has been investigated. The results demonstrate that E22K and R58Q, located in the immediate extension of the helices flanking the regulatory light chain Ca2+ binding site, had dramatically altered Ca2+ binding properties. The K Cavalue for E22K was decreased by ∼17-fold compared with the wild-type light chain, and the R58Q mutant did not bind Ca2+. Interestingly, Ca2+ binding to the R58Q mutant was restored upon phosphorylation, whereas the E22K mutant could not be phosphorylated. In addition, the α-helical content of phosphorylated R58Q greatly increased with Ca2+ binding. The A13T mutation, located near the phosphorylation site (Ser-15) of the human cardiac regulatory light chain, had 3-fold lowerK Ca than wild-type light chain, whereas phosphorylation of this mutant increased the Ca2+ affinity 6-fold. Whereas phosphorylation of wild-type light chain decreased its Ca2+ affinity, the opposite was true for A13T. The α-helical content of the A13T mutant returned to the level of wild-type light chain upon phosphorylation. The phosphorylation and Ca2+ binding properties of the regulatory light chain of human cardiac myosin are important for physiological function, and alteration any of these could contribute to the development of hypertrophic cardiomyopathy.
Intergenerational Programs in Schools

This study examined the prevalence, types, and perceived impact of intergenerational programs in schools. Programs involving senior volunteers assisting children, or children participating in activities with older people were considered. Of the schools reached, 47% reported intergenerational programming. Thirty-three schools in the Tel-Aviv region participated in the study. Data were collected from 85 seniors, 26 teachers, and 20 coordinators. Assessments included program characteristics, program preparation, and perceived benefits and difficulties. Both programs were reported to have beneficial effects for seniors and to benefit children in the academic, social, and emotional domains. However, programs appeared to attract different types of volunteers and different degrees of volunteer commitment. Findings suggest that there is a need to pay additional attention to both participants’ specific requests and needs and to the allocation of resources to improve the design and implementation of intergenerational programs.
Surface science investigations of the role of CO2 in astrophysical ices

We have recorded reflection–absorption infrared spectroscopy (RAIRS) and temperature-programmed desorption (TPD) data for a range of CO2-bearing model astrophysical ices adsorbed on a graphitic dust grain analogue surface. Data have been recorded for pure CO2, for CO2 adsorbed on top of amorphous solid water, for mixed CO2:H2O ices and for CO2 adsorbed on top of a mixed CH3OH:H2O ice. For the TPD data, kinetic parameters for desorption have been determined, and the trapping behaviour of the CO2 in the H2O (CH3OH) ice has been determined. Data of these types are important as they can be used to model desorption in a range of astrophysical environments. RAIR spectra have also shown the interaction of the CO2 with H2O and CH3OH and can be used to compare with astronomical observations, allowing the accurate assignment of spectra.
Distributions by Age and Sex of the Dimensions of Temperament and Character Inventory in a Cross-Cultural Perspective among Sweden, Germany, and the USA

Distribution by age and sex of the dimensions of the Temperament and Character Inventory were assessed cross-culturally for samples in Sweden, Germany, and the USA. The Temperament and Character Inventory is a 240-item (Sweden, 238-item), self-administered, true-false format, paper-and-pencil test developed by Cloninger and his coworkers based on his unified biosocial theory of personality. The inventory measures the Temperament dimensions Novelty Seeking, Harm Avoidance, Reward Dependence, and Persistence as well as the Character dimensions, Self-directedness, Cooperativeness, and Self-transcendence. The samples consisted of 300 German subjects, 300 Swedish subjects, and 300 U.S. subjects matched by age cohort and sex. Stability of the personality dimensions was evaluated across samples as were their age and sex distributions. We found significant effects of age, sex, and culture in univeriate and multivariate comparisons on the personality dimensions. However, several significant differences in the personality dimensions for both European samples appear to be similar compared with those of the U.S. sample. We have to conclude that sex- and age-specific norms for the dimensions of the Temperament and Character Inventory are necessary given the established significant differences.
Cellular and Molecular Targets of Benzo[a]pyrene and Metal Toxicity in Xenopus laevis Embryos and in Hep G2 Cells

This paper describes the use of two in vitro systems (stage 35 Xenopus laevis embryos and the human hepatoblastoma cell line, Hep G2) to study effects of some environmental contaminants (benzo[a]pyrene, copper and zinc), which are representative of chemicals with different cell targets and mechanisms of action. The ability to activate benzo[a]pyrene and to metabolise it with the cytochrome P4501A isozyme were demonstrated in both in vitro systems by assessing the formation of water-soluble and protein-bound benzo[a]pyrene metabolites and by immunochemical analysis. In X. laevis embryos, the formation of DNA adducts demonstrated the ability to produce benzo[a]pyrene reactive metabolites. Moreover, in Hep G2 cells, the cytoskeletal protein, tubulin, and the reduced form of glutathione proved to be the cellular targets of copper and zinc toxicity. In response to metal-induced stress in Hep G2 cells, there was a cytoplasmic reorganisation of heat shock protein, Hsp 70. In conclusion, the in vitro systems provide for a rapid evaluation of heterogeneous compounds such as benzo[a]pyrene and heavy metals that differ in toxic potency and mechanisms of action. They could also be used to study the mechanisms of toxic action and to identify specific cellular and molecular targets.
Neonatal skin care: clinical outcomes of the AWHONN/NANN evidence-based clinical practice guideline. Association of Women's Health, Obstetric and Neonatal Nurses and the National Association of Neonatal Nurses.

OBJECTIVE To test the effectiveness of an evidence-based clinical practice guideline for neonatal skin care on selected clinical outcomes for newborns in neonatal intensive-care units (NICU), special-care units (SCU), and well-baby nurseries. DESIGN Prospective evaluation of the collaborative neonatal skin care research-based practice project of the Association of Women's Health, Obstetric and Neonatal Nurses and the National Association of Neonatal Nurses. SETTING NICU and well-baby units in 51 hospitals located throughout the United States. PARTICIPANTS Member site coordinators (N = 51) and the neonates (N= 2,820) observed during both the pre- and postimplementation phases of the project. METHOD Site coordinators received specialized education in neonatal skin care and implemented an evidence-based clinical practice guideline addressing 10 aspects of neonatal skin care. Baseline observations of skin condition, care practices, and environment of newly admitted neonates were collected by site coordinators. Postimplementation observations were then completed. MAIN OUTCOME MEASURES Skin condition was assessed with the Neonatal Skin Condition Score (NSCS), which ranges from a score of three (best condition) to a score of nine (worst condition), based on dryness, erythema, and skin breakdown. Changes in frequency of selected skin care practices were used to assess the effectiveness and feasibility of using the practice guideline in everyday clinical practice. Aspects of the care environment with potential effect on skin integrity were monitored to determine risk factors. RESULTS Fifty-one site coordinators made 11,468 systematic assessments of 2,464 NICU and SCU newborns and 356 well newborns. Baseline skin scores were better in well newborns compared with premature newborns. After implementation of the guideline, skin condition was improved, as reflected by less visible dryness, redness, and skin breakdown in both the NICU/SCU and well newborns. The guideline was integrated into care, as evidenced by increased use of emollients, particularly with premature infants, and decreased frequency of bathing. A relationship was shown between selected aspects of the environment and alterations in skin integrity. CONCLUSIONS Use of the AWHONN/NANN Neonatal Skin Care Research-Based Clinical Practice Guideline was successfully implemented at 51 sites, and effectiveness was demonstrated by changed care practices and improved skin condition in premature and full-term newborns. The results of this project support a wider dissemination of the project's practice guideline for neonatal skin care.
Maturation of fetal hepatocytes in vitro by extracellular matrices and oncostatin M: Induction of tryptophan oxygenase

Previously, we described that embryonic day 14.5 (E14.5) mouse fetal hepatocytes differentiate to express tyrosine amino transferase (TAT) and glucose‐6‐phosphatase, which are expressed in the perinatal liver, in response to oncostatin M (OSM) or in high‐cell‐density culture. However, under such conditions, fetal hepatic cells failed to express genes for adult liver‐specific enzymes, such as tryptophan oxygenase (TO). Although phenobarbital (PB) and dimethylsulfoxide (DMSO) have been known to maintain the functions of adult hepatocytes in vitro, they failed to induce TO expression in fetal hepatic cells. Thus far, no system has been developed that reproduces terminal differentiation of fetal hepatocytes in vitro. Here, we describe that extracellular matrices derived from Engelbreth‐Holm‐Swarm sarcoma (EHS) in combination with OSM or high‐cell‐density culture induced expression of TO as well as cytochrome P450 genes that are involved in detoxification. However, EHS alone was insufficient to induce expression of TO, although it induced TAT expression in fetal hepatocytes. In addition, high‐density culture further augmented differentiation. In conclusion, the combination of signals by cytokines, cell‐cell contact, and cell‐matrix interaction is required for induction of adult liver functions in fetal hepatocytes in vitro. This primary culture system will be useful for studying the mechanism of liver development.
Low microsatellite instability is associated with poor prognosis in stage C colon cancer.

PURPOSE The significance of low microsatellite instability (MSI-L) in colorectal cancer is poorly understood. No clear biologic distinction has been found between MSI-L and microsatellite stable (MSS) colorectal cancer, and these two phenotypes are usually combined when analyzed against the well-defined high MSI (MSI-H) phenotype. Evidence is emerging that an O(6)-methylguanine DNA methyltransferase (MGMT) gene defect is associated with MSI-L. Therefore, to further define this phenotype, we undertook a detailed analysis of the prognostic significance of MSI-L and loss of MGMT expression in colon cancer. PATIENTS AND METHODS The study cohort was 183 patients with clinicopathologic stage C colon cancer who had not received adjuvant therapy. We analyzed MSI status, MGMT, and mismatch repair protein expression, as well as MGMT and p16 promoter hypermethylation. RESULTS We showed that MSI-L defines a group of patients with poorer survival (P = .026) than MSS patients, and that MSI-L was an independent prognostic indicator (P = .005) in stage C colon cancer. Loss of MGMT protein expression was associated with the MSI-L phenotype but was not a prognostic factor for overall survival in colon cancer. p16 methylation was significantly less frequent in MSI-L than in MSI-H and MSS tumors and was not associated with survival. CONCLUSION MSI-L characterizes a distinct subgroup of stage C colon cancer patients, including the MSI-L subset of proximal colon cancer, who have a poorer outcome. Neither the MGMT defect nor p16 methylation are likely to contribute to the worse prognosis of the MSI-L phenotype.
Early and late rate of force development: differential adaptive responses to resistance training?

The objective of this study is to investigate the potentially opposing influence of qualitative and quantitative muscular adaptations in response to high‐intensity resistance training on contractile rate of force development (RFD) in the early (<100 ms) and later phases (>200 ms) of rising muscle force. Fifteen healthy young males participated in a 14‐week resistance training intervention for the lower body and 10 matched subjects participated as controls. Maximal muscle strength (MVC) and RFD were measured during maximal voluntary isometric contraction of the quadriceps femoris muscle. Muscle biopsies were obtained from the vastus lateralis. The main findings were that RFD in the late phase of rising muscle force increased in response to resistance training whereas early RFD remained unchanged and early relative RFD (i.e., RFD/MVC) decreased. Quantitatively, muscle fiber cross‐sectional area and MVC increased whereas, qualitatively, the relative proportion of type IIX muscle fibers decreased. Multiple regression analysis showed that while increased MVC positively influenced both early and late RFD, decreased‐type IIX negatively influenced early RFD only. In conclusion, early and late RFD responded differently to high‐intensity resistance training due to differential influences of qualitative and quantitative muscular adaptations on early and later phases of rising muscle force.
Expression of the pS2 gene in normal, benign and neoplastic human stomach

The expression of the estrogenregulated breastcancerassociated pS2 gene was examined in 75 stomach resections taken from 45 patients. The 600‐base pS2 mRNA was found in all of the 47 nonneoplastic samples at varying levels: in the histologically normal group we observed a Poissontype distribution, whereas 79% of the tissues exhibiting dysplastic features expressed high levels of transcript. Tumour samples expressed relatively lower pS2 mRNA, with only 18% having high levels and 43% with no detectable expression. These differences were not correlated to tumour grading, stage or site. No amplification or rearrangement of the pS2 gene was found. Immunohistochemical analysis of formalinfixed paraffin sections, using a polyclonal antibody against pS2 protein, showed specific staining of both cytoplasm and membrane of epithelial cells in the neck region of antral and body glands as well as in luminal secretions. Immunoreactivity was observed in the subnuclear region of foveolar cells, with specialized gland and goblet cells in atrophic gastritis being negative. Heterogeneous but strong focal cytoplasmic staining was seen in tumour cells as well as in dysplastic epithelium. Two gastric cell lines, KATO III and MKN‐45, derived from poorly differentiated adenocarcinomas also expressed pS2, whereas 3 other lines from well differentiated parental tumours did not. Genomic analysis revealed a BamHI polymorphism in Kato III cells and in the nonexpressing MKN‐28 cells. Immunostaining to pS2 protein was also demonstrated in the cytoplasm of KATO III cells, but neither these nor any of 30 tissues examined showed any positivity with a monoclonal antibody (MAb) to estrogen receptor. Our results suggest that pS2 is normally expressed in human stomach, possibly in association with secretory activity, and becomes downregulated during malignancy.
Different fates of oocytes with DNA double-strand breaks in vitro and in vivo

In female mice, despite the presence of slight DNA double-strand breaks (DSBs), fully grown oocytes are able to undergo meiosis resumption as indicated by germinal vesicle breakdown (GVBD); however, severe DNA DSBs do reduce and delay entry into M phase through activation of the DNA damage checkpoint. But little is known about the effect of severe DNA DSBs on the spindle assembly checkpoint (SAC) during oocyte maturation. We showed that nearly no first polar body (PB1) was extruded at 12 h of in vitro maturation (IVM) in severe DNA DSBs oocytes, and the limited number of oocytes with PB1 were actually at telophase. However, about 60% of the severe DNA DSBs oocytes which underwent GVBD at 2 h of IVM released a PB1 at 18 h of IVM and these oocytes did reach the second metaphase (MII) stage. Chromosome spread at MI and MII stages showed that chromosomes fragmented after GVBD in severe DNA DSBs oocytes. The delayed PB1 extrusion was due to the disrupted attachment of microtubules to kinetochores and activation of the SAC. At the same time, misaligned chromosome fragments became obvious at the first metaphase (MI) in severe DNA DSBs oocytes. These data implied that the inactivation of SAC during the metaphase-anaphase transition of first meiosis was independent of chromosome integrity. Next, we induced DNA DSBs in vivo, and found that the number of superovulated oocytes per mouse was significantly reduced; moreover, this treatment increased the percentage of apoptotic oocytes. These results suggest that DNA DSBs oocytes undergo apoptosis in vivo.
TrailSense

Trail surface information is critical in preventing from the mountain accidents such as falls and slips. In this paper, we propose a new mobile crowdsensing system that automatically infers whether trail segments are risky to climb by using sensor data collected from multiple hikers’ smartphones. We extract cyclic gait-based features from walking motion data to train machine learning models, and multiple hikers’ results are then aggregated for robust classification. We evaluate our system with two real-world datasets. First, we collected data from 14 climbers for a mountain trail which includes 13 risky segments. The average accuracy of individuals is approximately 80%, but after clustering the results, our system can accurately identify all the risky segments. We then collected an additional dataset from five climbers in two different mountain trails, which have 10 risky segments in total. Our results show that the model trained in one trail can be used to accurately identify all the risky segments in the other trail, which documents the generalizability of our system.
Small molecule inhibitors of Wnt/beta-catenin/lef-1 signaling induces apoptosis in chronic lymphocytic leukemia cells in vitro and in vivo.

BACKGROUND Lymphoid enhancer factor-1 (lef-1) is overexpressed in B-cell chronic lymphocytic leukemia (CLL) when compared with normal B cells and transcribes several genes implicated in the pathogenesis of CLL. We therefore hypothesize that antagonism of lef-1 might lead to killing of CLL cells. We used two small molecule inhibitors of Wnt/beta-catenin/lef-1 signaling (CGP049090 and PKF115-584) to test our hypothesis. DESIGN AND METHODS Enriched CLL cells and healthy B cells were used in this study. Small interfering RNA (siRNA)-mediated knockdown of lef-1 in primary CLL cells was done using nucleofection, and 50% lethal concentration (LC(50)) of two small molecules was assessed using ATP-based cell viability assay. Apoptotic response was investigated in time course experiments with different apoptotic markers. Specificity of the small molecules was demonstrated by coimmunoprecipitation experiments for the lef-1/beta-catenin interaction. In vivo studies were done in JVM-3 subcutaneous xenograft model. RESULTS Inhibition of lef-1 by siRNA leads to increased apoptosis of CLL cells and inhibited proliferation of JVM-3 cell lines. The two small molecule inhibitors (CGP049090 and PKF115-584) efficiently kill CLL cells (LC(50)<1 microM), whereas normal B cells were not significantly affected. Coimmunoprecipitation showed a selective disruption of beta-catenin/lef-1 interaction. In vivo studies exhibited tumor inhibition of 69% with CGP049090 and 57% with PKF115-584 when compared with vehicle-treated controls, and the intervention was well tolerated. CONCLUSIONS We have demonstrated that targeting lef-1 is a new and selective therapeutic approach in CLL. CGP049090 or PKF115-584 may be attractive compounds for CLL and other malignancies that deserve further (pre)clinical evaluation.
Regulation of the IL-10-driven macrophage phenotype under incoherent stimuli

Macrophages are ubiquitous innate immune cells that play a central role in health and disease by adopting distinct phenotypes, which are broadly divided into classical inflammatory responses and alternative responses that promote immune suppression and wound healing. Although macrophages are attractive therapeutic targets, incomplete understanding of this functional choice limits clinical manipulation. While individual stimuli, pathways, and genes involved in macrophage functional responses have been identified, how macrophages evaluate complex in vivo milieus comprising multiple divergent stimuli remains poorly understood. Here, we used combinations of “incoherent” stimuli—those that individually promote distinct macrophage phenotypes—to elucidate how the immunosuppressive, IL-10-driven macrophage phenotype is induced, maintained, and modulated under such combinatorial stimuli. The IL-10-induced immunosuppressive phenotype was largely insensitive to co-administered IL-12, which has been reported to modulate macrophage phenotype, but maintaining the immunosuppressive phenotype required sustained exposure to IL-10. Our data implicate the intracellular protein, BCL3, as a key mediator of the IL-10-driven phenotype. Notably, co-administration of IFN-γ disrupted an IL-10-mediated positive feedback loop that may reinforce the immunosuppressive phenotype. This novel combinatorial perturbation approach thus generated new insights into macrophage decision making and local immune network function.
Characterization and Multipotentiality of Human Fetal Femur–Derived Cells: Implications for Skeletal Tissue Regeneration

To date, the plasticity, multipotentiality, and characteristics of progenitor cells from fetal skeletal tissue remain poorly defined. This study has examined cell populations from human fetal femurs in comparison with adult‐derived mesenchymal cell populations. Real‐time quantitative polymerase chain reaction demonstrated expression of mesenchymal progenitor cell markers by fetal‐derived cells in comparison with unselected adult‐derived and immunoselected STRO‐1–enriched adult populations. Multipotentiality was examined using cells derived from femurs and single‐cell clones, culture‐expanded from explants, and maintained in basal medium prior to exposure to adipogenic, osteogenic, and chondrogenic conditions. Adipocyte formation was confirmed by Oil Red O lipid staining and aP2 immunocytochemistry, with expression of peroxisome proliferation‐activated receptor‐γ detected only in adipogenic conditions. In chondrogenic pellets, chondrocytes lodged within lacunae and embedded within dense proteoglycan matrix were observed using Alcian blue/Sirius red staining and type II collagen immunocytochemistry. Osteogenic differentiation was confirmed by alkaline phosphatase staining and type I collagen immunocytochemistry as well as by gene expression of osteopontin and osteocalcin. Single‐cell clonal analysis was used to demonstrate multipotentiality of the fetal‐derived populations with the formation of adipogenic, chondrogenic, and osteogenic populations. Mineralization and osteoid formation were observed after culture on biomimetic scaffolds with extensive matrix accumulation both in vitro and in vivo after subcutaneous implantation in severely compromised immunodeficient mice. These studies demonstrate the proliferative and multipotential properties of fetal femur–derived cells in comparison with adult‐derived cells. Selective differentiation and immunophenotyping will determine the potential of these fetal cells as a unique alternative model and cell source in the restoration of damaged tissue.