Miracle-dz/UniFAR-FTUs huggingface dataset:
Modified yeast cells to investigate the coupling of G protein‐coupled receptors to specific G proteins

G protein‐coupled receptors (GPCRs) help to regulate the physiology of all the major organ systems. They respond to a multitude of ligands and activate a range of effector proteins to bring about the appropriate cellular response. The choice of effector is largely determined by the interaction of individual GPCRs with different G proteins. Several factors influence this interaction, and a better understanding of the process may enable a more rational approach to identifying compounds that affect particular signalling pathways. A number of systems have been developed for the analysis of GPCRs. All provide useful information, but the genetic amenability and relative simplicity of yeast makes them a particularly attractive option for ligand identification and pharmaceutical screening. Many, but not all, GPCRs are functional in the budding yeast Saccharomyces cerevisiae, and we have developed reporter strains of the fission yeast Schizosaccharomyces pombe as an alternative host. To provide a more generic system for investigating GPCRs, we created a series of yeast–human Gα‐transplants, in which the last five residues at the C‐terminus of the yeast Gα‐subunit are replaced with the corresponding residues from different human G proteins. These enable GPCRs to be coupled to the Sz. pombe signalling machinery so that stimulation with an appropriate ligand induces the expression of a signal‐dependent lacZ reporter gene. We demonstrate the specificity of the system using corticotropin releasing factor (CRF) and CRF‐related peptides on two CRF receptors. We find that different combinations of ligand and receptor activate different Gα‐transplants, and the specificity of the coupling is similar to that in mammalian systems. Thus, CRF signalled through the Gs‐ and Gi‐transplants, consistent with its regulation of adenylate cyclase, and was more active against the CRF‐R1A receptor than against the CRF‐R2B receptor. In contrast, urocortin II and urocortin III were selective for the CRF‐R2B receptors. Furthermore, urocortin, but not CRF, induced signalling through the CRF‐R1A receptor and the Gq‐transplant. This is the first time that human GPCRs have been coupled to the signalling pathway in Sz. pombe, and the strains described in this study will complement the other systems available for studying this important family of receptors.
Basement membrane composition in the early mouse embryo day 7

Basement membranes (BM) are specialized structures of the extracellular matrix known to be involved in various early developmental processes. Despite numerous investigations on the localization of BM components, it remains unknown which molecules are expressed in early developmental stages and by which germ layers these proteins are produced. Therefore, we tested for all known laminin chains, nidogens, collagen type IV, and perlecan by means of light microscopic immunostaining and performed in situ reverse transcriptase‐polymerase chain reaction to detect the mRNAs specific for laminin α1, laminin β1, the α1 chain of collagen type IV, nidogen‐2, and perlecan in the early mouse embryo, day 7, in vivo. Only the laminin chains α1, β1, and γ1 were detected immunohistochemically throughout the entire endodermal and ectodermal BM zones of the embryo proper. The mRNA of laminin α1, laminin β1, collagen type IV, nidogen‐2 and perlecan were expressed in the ectoderm‐derived mesoderm, in the endoderm as well as in the ectoderm. In contrast, Reichert's membrane was positive for all laminin chains except for the α4, α5, β3, and γ3 chains. Moreover, maternal epithelial as well as mesenchymal cells expressed laminins, nidogen‐1 and nidogen‐2, collagen type IV, and perlecan. In conclusion, laminin‐1 might be the only laminin isoform in the early mouse embryo that, together with the other main BM components, nidogens, collagen type IV, and perlecan, is synthesized by all three germ layers. Developmental Dynamics 233:1140–1148, 2005. © 2005 Wiley‐Liss, Inc.
Terminal illness: views of patients and their lay carers

This paper examines the responses of 28 terminally ill patients and their lay carers to identical questions about the patients' experiences in the month prior to their admission to an inpatient hospice. No statistically significant differences were found between the reports of patients or carers regarding the patients' activities of daily living, their physical symptoms, or the evaluations of the care they received. Discrepancies between accounts were most likely to concern the presence or absence of psychological symptoms, the degree of distress caused to patients by their symptoms, and the main symptoms experienced by the patient on their admission to the hospice. It is suggested that the accounts of patients' experiences given by their carers can be used as a valid source of information.
The effect of the price or rental cost of housing on family size: a theoretical analysis with reference to New Zealand

ABSTRACT We investigate the effect of higher house prices or rental costs on family size. We provide a static model of a household's choice of family size assuming constant elasticity of substitution preferences between children, leisure, and other goods, and Cobb–Douglas household production of children using parental time and housing. We then explore the wealth and substitution effects that changing house prices or rental costs have on desired family size. Renters are predicted to have fewer children in response to higher rents. Home owners are predicted to have more children in response to higher house prices only if they have sufficient initial housing and low substitution between family size and other consumption, and fewer children otherwise. Finally, we provide exploratory correlations between the change in lagged house prices or rental costs and the change in number of children born to women using aggregated census units in New Zealand. We find weak negative correlations in both cases.
Late ureteral obstruction after kidney transplantation

Today, the incidence of urological complications following renal transplantation is 2%–10%. Most of these complications occur within the 1st year and affect the distal ureter. We report on two patients who developed very late ureteral obstruction, 14 and 18 years after transplantation. Both patients had rejection episodes 1 and 10 months prior to the ureteral stenosis. Histological examination of one resected ureter revealed findings strongly suggestive of a rejection process. Open surgery with antirefluxive reimplantation into the bladder was successful in both patients, with a postoperative observation time of 20 and 8 months, respectively. We conclude that a percutaneous nephrostomy may be required in patients with rising creatinine and incipient hydronephrosis even long after transplantation has been performed.
Coping and psychological adjustment in adolescents with vertically acquired HIV

Abstract Children with vertically acquired HIV face the challenges of adolescence in addition to the demands of coping with their illness. The relationship between coping and psychological adjustment has been widely studied in adults and children with chronic diseases but it is poorly understood in adolescents with HIV. This study aimed to identify whether a UK sample of adolescents with vertically acquired HIV had poor psychological adjustment and to clarify the relationship between coping and psychological adjustment in this sample. Thirty adolescents with vertically acquired HIV (aged 11–17) filled in questionnaires of coping and psychological adjustment and a correlational design was used to determine if specific coping styles were related to quality of psychological adjustment. Results showed that younger children had lower levels of psychological adjustment than older adolescents, although as a group the adolescents scored within normal ranges. Psychological adjustment was significantly related to two coping styles, Ventilating feelings and Being humorous. Exploratory analysis examined the extent of HIV disclosure in this sample and the degree of satisfaction felt about the nature of communication about the diagnosis. The results showed that participants had few confidants but were satisfied with the nature of communication about their status. The level of satisfaction was not related to adjustment. On the basis of the results, potential clinical implications for adolescents with vertically acquired HIV who might be struggling not only with the challenging demands of adolescence but also with their illness are discussed. The need to adopt a developmental approach to coping and psychological adjustment is emphasized.
Characterization of Recombinant Soluble Macrophage Scavenger Receptor MARCO*

MARCO is a type II transmembrane protein of the class A scavenger receptor family. It has a short N-terminal cytoplasmic domain, a transmembrane domain, and a large extracellular part composed of a 75-residue long spacer domain, a 270-residue collagenous domain, and a 99-residue long scavenger receptor cysteine-rich (SRCR) domain. Previous studies have indicated a role for this receptor in anti-microbial host defense functions. In this work we have produced the extracellular part of MARCO as a recombinant protein, and analyzed its binding properties. The production of this protein, soluble MARCO (sMARCO), has made it possible for the first time to study MARCO and its binding properties in a cell-free system. Using circular dichroism analyses, a protease-sensitive assay, and rotary shadowing electron microscopy, sMARCO was shown to have a triple-helical collagenous structure. Rotary shadowing also demonstrated that the molecules often associate with each other via the globes. sMARCO was found to bind avidly both heat-killed and living bacteria. Lipopolysaccharide, an important component of the outer membrane of Gram-negative bacteria, was shown to be a ligand of MARCO. Studies with different bacterial strains indicated that the O-side chain of lipopolysaccharide is not needed for the bacterial recognition. Finally, the C-terminal SRCR domain was also produced as a recombinant protein, and its bacteria-binding capability was studied. Although the transfection experiments with transmembrane MARCO variants have indicated a crucial role for this domain in bacterial binding, the monomeric domain exhibited low, barely detectable bacteria-binding activity. Thus, it is possible that cooperation between the SRCR domain and the collagenous domain is needed for high-affinity bacterial binding, or that the SRCR domain has to be in a trimeric form to effectively bind to bacteria.
Sensor fusion and computer vision for context-aware control of a multi degree-of-freedom prosthesis

Objective. Myoelectric activity volitionally generated by the user is often used for controlling hand prostheses in order to replicate the synergistic actions of muscles in healthy humans during grasping. Muscle synergies in healthy humans are based on the integration of visual perception, heuristics and proprioception. Here, we demonstrate how sensor fusion that combines artificial vision and proprioceptive information with the high-level processing characteristics of biological systems can be effectively used in transradial prosthesis control. Approach. We developed a novel context- and user-aware prosthesis (CASP) controller integrating computer vision and inertial sensing with myoelectric activity in order to achieve semi-autonomous and reactive control of a prosthetic hand. The presented method semi-automatically provides simultaneous and proportional control of multiple degrees-of-freedom (DOFs), thus decreasing overall physical effort while retaining full user control. The system was compared against the major commercial state-of-the art myoelectric control system in ten able-bodied and one amputee subject. All subjects used transradial prosthesis with an active wrist to grasp objects typically associated with activities of daily living. Main results. The CASP significantly outperformed the myoelectric interface when controlling all of the prosthesis DOF. However, when tested with less complex prosthetic system (smaller number of DOF), the CASP was slower but resulted with reaching motions that contained less compensatory movements. Another important finding is that the CASP system required minimal user adaptation and training. Significance. The CASP constitutes a substantial improvement for the control of multi-DOF prostheses. The application of the CASP will have a significant impact when translated to real-life scenarious, particularly with respect to improving the usability and acceptance of highly complex systems (e.g., full prosthetic arms) by amputees.
Cholesterol is Required for the Formation of Regulated and Constitutive Secretory Vesicles from the trans‐Golgi Network

We studied the role of cholesterol in regulated protein secretion in neuroendocrine cells by manipulating the cholesterol content of AtT‐20 cells. Depletion of cellular cholesterol levels caused a reversible block of immature secretory granule biogenesis at the level of the trans‐Golgi‐network, whereas increased cholesterol levels promoted immature secretory granule formation. Cholesterol depletion also blocked the formation of constitutive secretory vesicles, but did not inhibit the transport between the endoplasmic reticulum and the Golgi complex. Our results indicate that the assembly of cholesterol‐based lipid microdomains is required for the biogenesis of both regulated and constitutive secretory vesicles from the trans‐Golgi‐network in neuroendocrine cells.
FUNCTION OF THE TRANSCRIPTIONAL REGULATING PROTEIN OF 132 kDa (TReP-132) ON HUMAN P450scc GENE EXPRESSION

Cytochrome P450scc catalyzes the important first step in the steroid synthesis pathway; however, it is clear that additional factors regulating the temporal and spacial specific expression of the CYP11A1 gene remain to be identified. To isolate novel transcription factors that regulate this gene, a cis-acting element of the 5′-flanking region from nucleotides −155 to −131 (−155/−131) was used to screen a human placental λgt11 cDNA expression library, and an interacting clone was isolated. The open reading frame of the cDNA encodes several domains that are characteristic of transcription factors including an acidic region, a region rich in prolines and three zinc-finger motifs. Expression of the cDNA by in vitro transcription/translation and by transient transfection in HeLa cells yielded a protein of 132 kDa, which concurs with the predicted size. Transfection of the cDNA in placental JEG-3 and adrenal NCI-H295 cells, stimulate expression of a reporter construct controlled by the P450scc gene 5′-flanking region from nucleotides −1676 to +49. This transcriptional regulating protein of 132 kDa (TReP-132) when expressed in HeLa cells was demonstrated to interact with the −155/−131 region in bandshift analysis, and tandem copies of this region was shown to confer activation of the heterologous HSV thymidine kinase minimal promoter. Coexpression of CBP/p300 with TReP-132 further increased promoter activity, and the proteins were demonstrated to interact physically. RNA analysis demonstrated the highest levels of expression in the adrenal cortex and testis; and transcript expression is also found in the steroidogenic JEG-3, NCI-H295, and MCF-7 cell lines, but not in non-steroidogenic HepG2 and HK293 cells. Subsequently it has been shown that TReP-132 interacts with steroidogenic factor-1 (SF-1) through specific domains; and along with the interaction with CBP/p300 these factors are postulated to form a complex to regulate expression of the P450scc gene.
Lack of Association between Air Pollutant Exposure and Short-Term Risk of Ischaemic Stroke in Lyon, France

Background Some observational and experimental studies have suggested a short-term relationship between air pollutants and ischaemic stroke; however, the results conflict. Aims The objective of this study was to investigate the association between particulate matter less than 2·5 and 10 microns in aerodynamic diameter, nitrogen dioxide, sulphur dioxide and ozone, and short-term risk of ischaemic stroke in Lyon, France. Methods The AVC69 study was a multicenter cohort study in which all consecutive adult patients admitted to one of the emergency or neurological departments of the Rhone area for suspicion of stroke were included during a seven-month period. Only patients with ischaemic stroke living within the study area, composed of Lyon and 18 neighbouring communities with homogenous air pollutants exposure, formed the basis of our study. We adopted a time-stratified case-crossover design to analyse the short-term effect (up to two-days) of air pollutants on ischaemic stroke incidence. Models were adjusted for temperature, variation of atmospheric pressure, minimal relative humidity, influenza epidemics, pollen count, and holidays. Stratified analyses by gender and class age were performed. Different lag times were analysed. Results 376 patients were included. Mean age was 76·6 years (±13·7). 53·7% were women. No association was observed between air pollutants and short-term risk of ischaemic stroke after adjustment for main confounding factors. Results remained unchanged whatever the gender or age. Conclusions These results suggest a lack of association between air pollutant exposure and short-term risk of ischaemic stroke in a French urban area.
New product and service launching in new ventures: a multilevel approach to innovation antecedents

ABSTRACT Macro-economic policies such as public financing seek to push the development and introduction of innovations; however, entrepreneurs also need support in their ‘day-to-day’ activities to improve their capability to launch innovations. As this micro-level perspective is rarely studied, we analyze both the micro and macro levels by examining the effects of the entrepreneurs’ individual intention to innovate and public financing. Additionally, we include the meso level, representing entrepreneurs’ network. Entrepreneurs are embedded in social spheres in which they capture resources and identify opportunities. But not all entrepreneurs are equally well supported, and some tend to be completely isolated. We thus focus on multilevel factors explaining new product or service launching in new ventures. Drawing on data of 48,251 French new ventures, we reveal that innovation intention and public financing positively impact new ventures’ product and service innovation launching, while entrepreneurs’ isolation has a negative effect. We also highlight two interaction effects that enhance the multilevel effects of innovation antecedents.
Global and non‐random CpG‐island methylation in gastric carcinoma associated with Epstein‐Barr virus

DNA hypermethylation may play a primary role in the genesis of Epstein‐Barr virus (EBV)‐associated gastric carcinoma (GC) (EB‐VaGC). Methylation‐specific PCR targeting CpG‐islands demonstrated markedly increased methylation of specific genes, such as p14, p15 and p16 genes, in EBVaGC in vivo. A high frequency of methylation was observed in an EBVaGC strain of severe combined immunodeficiency mice, and the expression of methylated genes in the strain was apparently lower than the expression of the unmethylated genes in EBV‐negative GC strains. Although over‐expression of DNA methyltransferases (DNMTs) is known to be associated with some human cancers, real‐time PCR demonstrated that DNMTs expression was suppressed in EBVaGC. The DNA methylation of specific genes, independently of DNMTs expression, may be important in the development of EBVaGC. (Cancer Sci 2003; 94: 76–80)
Probabilistic small signal stability analysis of power system with large scale wind power

Large-scale new energy grid brings plenty of influence to the power system, so research on wind turbine grid for power system stability has become a hot topic. Although the traditional small signal stability analysis method of the deterministic power system is relatively mature, the uncertainty of wind turbine output brings challenge to the stability analysis of the power system connected with large-scale wind turbines. The discrete point estimation method which is also the uncertain approximation theory is applied into the stability analysis of the power system involved a massive wind turbines in this paper, based on the traditional small signal stability analysis of deterministic power system. In order to verify the correctness of the method proposed in this paper, experiments of the New England 39 node 10 machine power system based on Monte Carlo simulation (MCS) were performed.
Deoxyribonucleic Acid Relatedness Among Species of Erwinia and Between Erwinia Species and Other Enterobacteria

Relatedness in species of Erwinia was assessed by determining the extent of reassociation in heterologous deoxyribonucleic acid preparations. Thermal elution chromatography on hydroxyapatite was used to separate reassociated nucleotide sequences from nonreassociated sequences and to determine the thermal stability of related nucleotide sequences. An apparent 15% core of relatedness is present between fire blight, soft-rot, and “atypical” Erwinia species. All Erwinia species showed low to moderate reaction with representative enteric bacteria.
Are cardiorespiratory fitness and walking performance associated with self-reported quality of life and work ability?

OBJECTIVES In Finland, testing cardiorespiratory fitness (VO2max) is popular in health promotion programs and work ability evaluations. The most common instruments used for this purpose are the submaximal cycle ergometer test, and the 2-km walking test. However, limited data exist on the associations between VO2max and wellbeing in working age adults. The aim of the study was to evaluate how the measured (cycle ergometer) and the estimated (walking test) VO2max and walking performance are associated with health-related quality of life and work ability. MATERIALS AND METHODS The subjects were 104 middle-aged men workers (45-55 years old), mostly from the construction and manufacturing industries. VO2max was directly measured by a maximal exercise test on a cycle ergometer. The 2-km walking test parameters were the walking time, predicted VO2max, and fitness index. Health-related quality of life was assessed with the RAND-36 questionnaire which was further divided into physical dimensions (P-RAND-36) and mental dimensions (M-RAND-36). Perceived work ability was assessed with the work ability index (WAI) in a subgroup of 51 subjects. RESULTS The 2-km walking test parameters significantly predicted the score on P-RAND-36 (r2=0.18, p=0.001), and correlated significantly with WAI. The directly measured VO2max was not associated with P-RAND-36, M-RAND-36 or WAI. CONCLUSIONS The inexpensive 2-km walking test may be more useful when evaluating the quality of life and work ability, compared to the more expensive direct measurement of one's cardiorespiratory fitness in a laboratory.