Miracle-dz/UniFAR-FTUs huggingface dataset:
Expression of chimeric genes by the light-regulated cabII-1 promoter in Chlamydomonas reinhardtii: a cabII-1/nit1 gene functions as a dominant selectable marker in a nit1- nit2- strain

In Chlamydomonas reinhardtii, expression of the cabII-1 gene increases dramatically in response to light (cabII-1 encodes one of the light-harvesting chlorophyll a/b-binding proteins of photosystem II). We have used a region upstream of the cabII-1 gene in translational fusions to the bacterial uidA gene (encodes beta-glucuronidase) and transcriptional fusions to the Chlamydomonas nitrate reductase gene (nit1). Chlamydomonas transformants carrying intact copies of the chimeric uidA gene do not express beta-glucuronidase at the level of enzyme activity or mRNA accumulation. Methylation in the cabII-1 promoter region of the introduced gene is extensive in these strains, suggesting that newly introduced foreign genes may be recognized and silenced by a cellular mechanism that is correlated with increased methylation. Transformants that express the chimeric cabII-1/nit1 gene have been recovered. In contrast to the endogenous nit1 gene, the chimeric cabII-1/nit1 gene is expressed in ammonium-containing medium. Moreover, nit1 mRNA accumulation is dramatically stimulated by light, with a time course that is indistinguishable from that of the endogenous cabII-1 gene. The cabII-1/nit1 gene has been used to select transformants in a nit1- nit2- Chlamydomonas strain (CC400G) and should be useful for transformation of the large number of mutants in the Ebersold-Levine lineage, which carry the same mutations.
Early Pulmonary Complications following Total Knee Arthroplasty under General Anesthesia: A Prospective Cohort Study Using CT Scan

Purpose. Postoperative pulmonary complications (PPCs) are common after major surgeries. However, the number of studies regarding PPCs following total knee arthroplasty (TKA) is limited. The aim of this study was to determine the incidence of early PPCs following TKA by computed tomography (CT) scan and to identify associated risk factors. Methods. Patients, who were diagnosed with osteoarthritis or rheumatoid arthritis and underwent primary TKA at our institution, were included in this prospective cohort study. Patients received a standard procedure of TKA under general anesthesia. Chest CT scan was performed during 5–7 days postoperatively. Univariate analysis and multivariate logistic regression analysis were employed to identify the risk factors. Results. The total incidence of early PPCs following TKA was 45.9%. Rates of pneumonia, pleural effusion, and atelectasis were 14.4%, 38.7%, and 12.6%, respectively. Lower body mass index and perioperative blood transfusion were independent risk factors for PPCs as a whole and associated with atelectasis. Postoperative acute episode of hypoxemia increased the risk of pneumonia. Blood transfusion alone was related to pleural effusion. Conclusions. The incidence of early PPCs following TKA was high. For patients with relevant risk factors, positive measures should be adopted to prevent PPCs.
Electromagnetic Analysis of the Voltage-Temperature Characteristics of the ITER TF Conductor Samples

The measurement of the current sharing temperature of a Cable in Conduit Conductor is a complex task. The voltage traces measured on the conduit (jacket) of these conductors during tests, give V-I characteristics that significantly differ from the typical transition from the superconducting to the resistive state measured on single strands. In these measurements, after an initial ramping of the transport current up to the test value, the temperature is increased by steps until the resistive transition occurs. However, even in the first phase of the current ramping, when the cable is still fully or almost superconductive, early voltages are measured along the jacket at different angular positions around the cable. In particular, this was observed in the ITER TF conductors recently tested in the SULTAN facility at Villigen PSI, Switzerland. These samples showed significant ramps of voltages taken in the proximity of the joints and terminations. The present paper gives a possible qualitative and quantitative explanation for this mechanism. Numerical simulations are also shown which reproduce the scattering of the voltages measured around the jacket at a given location, showing the effect of the voltage taps position along the jacket in a six sub-cable model.
Wnt/beta-catenin signaling mediates oval cell response in rodents.

UNLABELLED Adult hepatic stem cells or oval cells are facultative stem cells in the liver that are activated during regeneration only during inhibition of innate hepatocyte proliferation. On the basis of its involvement in liver cancer, regeneration, and development, we investigated the role of the Wnt/beta-catenin pathway in oval cell response, which was initiated in male Fisher rats with 2-acetylaminofluorine and two-third partial hepatectomy (PHX). Extensive oval cell activation and proliferation were observed at 5 and 10 days post-PHX, as indicated by hematoxylin-eosin and proliferating cell nuclear antigen analysis. A noteworthy increase in total and active beta-catenin was observed at this time, which was localized to the oval cell cytoplasm and nuclei by immunohistochemistry and confirmed by double immunofluorescence. A concomitant increase in Wnt-1 in hepatocytes along with increased expression of Frizzled-2 in oval cells was observed. This paracrine mechanism coincided with a decrease in Wnt inhibitory factor-1 and glycogen synthase kinase-3beta down-regulation leading to beta-catenin stabilization. To strengthen its role, beta-catenin conditional knockout mice were treated with 3,5-diethoxycarbonyl-1,4-dihydrocollidine to induce oval cell activation. A dramatic decrease in the A6-positive oval cell numbers in the absence of beta-catenin demonstrated a critical role of beta-catenin in oval cell biology. CONCLUSION The Wnt/beta-catenin pathway plays a key role in the normal activation and proliferation of adult hepatic stem cells.
Anatomical variations of the Internal Jugular Vein in Non-Uremic Outpatients

The internal jugular vein (IJV) should be the preferred form of venous vascular access for the placement of dialysis catheters. “Blind” puncture or “skin mark” ultrasound technique puncture present multiple complications due to the significant variations in IJV location in normal subjects and even more so in uremic patients. The aim of this study is to demonstrate the important rate of IJV site variations in a random healthy population. We tested 450 subjects (244 male, 206 female) in our hospital Ultrasound Ambulatory using an Ansaldo AUS ultrasound device with linear 10 MHz probe on both sides at the Sedillot triangle level and noticed the relations between IJV and carotid artery course. The most frequent location of IJV was the anterior lateral (79.3% on the right side, 83.5% on the left). The remaining options were lateral, anterior, posterior-lateral, anterior-medial. We found no significant difference in IJV diameter on either side of the neck. We therefore consider IJV puncture with ultrasound guidance to be the first choice in central venous cannulation for hemodialysis treatment. This procedure helps avoid incorrect puncture of the carotid artery related to its abnormal location.
Effects of Ebola Virus Glycoproteins on Endothelial Cell Activation and Barrier Function

ABSTRACT Ebola virus causes severe hemorrhagic fever with high mortality rates in humans and nonhuman primates. Vascular instability and dysregulation are disease-decisive symptoms during severe infection. While the transmembrane glycoprotein GP1,2 has been shown to cause endothelial cell destruction, the role of the soluble glycoproteins in pathogenesis is largely unknown; however, they are hypothesized to be of biological relevance in terms of target cell activation and/or increase of endothelial permeability. Here we show that virus-like particles (VLPs) consisting of the Ebola virus matrix protein VP40 and GP1,2 were able to activate endothelial cells and induce a decrease in barrier function as determined by impedance spectroscopy and hydraulic conductivity measurements. In contrast, the soluble glycoproteins sGP and Δ-peptide did not activate endothelial cells or change the endothelial barrier function. The VLP-induced decrease in barrier function was further enhanced by the cytokine tumor necrosis factor alpha (TNF-α), which is known to induce a long-lasting decrease in endothelial cell barrier function and is hypothesized to play a key role in Ebola virus pathogenesis. Surprisingly, sGP, but not Δ-peptide, induced a recovery of endothelial barrier function following treatment with TNF-α. Our results demonstrate that Ebola virus GP1,2 in its particle-associated form mediates endothelial cell activation and a decrease in endothelial cell barrier function. Furthermore, sGP, the major soluble glycoprotein of Ebola virus, seems to possess an anti-inflammatory role by protecting the endothelial cell barrier function.
Measuring mechanical properties in cells: three easy methods for biologists

The mechanism by which cells sense stresses and transmit them throughout the cytoplasm and the cytoskeleton (CSK) and by which these mechanical signals are converted into biochemical signaling responses is not clear. Specifically, there is little direct experimental evidence on how intracellular CSK structural elements in living cells deform and transmit stresses in response to external mechanical forces. Existing theories have invoked various biophysical and biochemical mechanisms to explain how cells spread, deform, divide, move, and change shape in response to mechanical inputs, but rigorous tests in cells are lacking. The lack of data and understanding is preventing the identification of mechanisms and sites of mechano‐regulation in cells. Here, we introduce and describe three unique and easy methods for biologists to determine mechanical properties and signaling events in cells.
Neither Agouti-Related Protein nor Neuropeptide Y Is Critically Required for the Regulation of Energy Homeostasis in Mice

ABSTRACT Agouti-related protein (AgRP), a neuropeptide abundantly expressed in the arcuate nucleus of the hypothalamus, potently stimulates feeding and body weight gain in rodents. AgRP is believed to exert its effects through the blockade of signaling by α-melanocyte-stimulating hormone at central nervous system (CNS) melanocortin-3 receptor (Mc3r) and Mc4r. We generated AgRP-deficient (Agrp−/−) mice to examine the physiological role of AgRP. Agrp−/− mice are viable and exhibit normal locomotor activity, growth rates, body composition, and food intake. Additionally, Agrp−/− mice display normal responses to starvation, diet-induced obesity, and the administration of exogenous leptin or neuropeptide Y (NPY). In situ hybridization failed to detect altered CNS expression levels for proopiomelanocortin, Mc3r, Mc4r, or NPY mRNAs in Agrp−/− mice. As AgRP and the orexigenic peptide NPY are coexpressed in neurons of the arcuate nucleus, we generated AgRP and NPY double-knockout (Agrp−/−;Npy−/−) mice to determine whether NPY or AgRP plays a compensatory role in Agrp−/− or NPY-deficient (Npy−/−) mice, respectively. Similarly to mice deficient in either AgRP or NPY, Agrp−/−;Npy−/− mice suffer no obvious feeding or body weight deficits and maintain a normal response to starvation. Our results demonstrate that neither AgRP nor NPY is a critically required orexigenic factor, suggesting that other pathways capable of regulating energy homeostasis can compensate for the loss of both AgRP and NPY.
Sensitivity of IL‐5 production to the cAMP‐dependent pathway in human T cells is reduced by exogenous IL‐2 in a phosphoinositide 3‐kinase‐dependent way

The cAMP‐dependent pathway plays an important role in the regulation of T cell‐mediated immune responses by inhibition of T cell proliferation, activation and production of Th1‐like cytokines.Depending on costimulatory signals and on the activation status of T cells, cAMP also regulates the production of Th2‐like cytokines, yet the mechanism is not completely defined. We investigated the effect of costimulation with IL‐2 on cAMP‐mediated inhibition of IL‐5 secretion and the signaling pathways involved in these effects in freshly isolated, α‐CD3/α‐CD28‐stimulated human T lymphocytes. We demonstrate that IL‐2 counteracts the cAMP‐mediated inhibitory effects on IL‐5 secretion by the modulation of phosphoinositide 3‐kinase (PI3‐K)‐dependent signaling. Our results indicate that phosphorylation of cAMP‐responsive element‐binding protein (CREB) and the activity of the small GTPase Rap1 are unlikely involved in the protective effect of IL‐2. Instead, the effect of IL‐2may be mediated by the PI3‐K‐dependent inactivation of the forkhead‐related transcription factor FKHR‐L1, down‐regulation of p27kip and abrogation of the cAMP‐mediated inhibition of activator protein (AP)‐1 binding activity. Together, our results indicate that increased IL‐2‐dependent PI3‐K signaling leads to impaired negative feedback control of the production of Th2‐type cytokine IL‐5 by the cAMP‐dependent pathway.
Changes in Energy Intensity in Canada

Canada is one of the top energy users and CO2 emitters among the OECD countries. However, energy intensity has been declining, on average, by about 1.4 percent since 1980. In this paper, we use the Fisher Ideal Index to determine the contribution of changes in the composition of economic activities and efficiency to a decline in energy intensity in Canada at national, provincial, and industry levels. We also apply panel data estimation methods to further investigate the factors driving energy intensity, efficiency and activity indexes for the period 1981-2008. We test for endogeneity as well as cross-section dependency in the provincial data and control for factors such as climate, policy, and energy endowment. The national and provincial decomposition results suggest that most of the reduction in energy intensity has occurred mainly due to improvements in energy efficiency rather than shifts in economic activities. Within the industry, while energy intensity has declined significantly in manufacturing, it has remained stable in transportation, utilities, and construction, and increased significantly in oil extraction and mining industries. The provincial panel regression results indicate that energy intensity is higher in provinces with higher average incomes, faster population growth, colder climate, and a higher capital-labour ratio, and lower in provinces with higher energy prices and higher investment. The industry panel regression results show that investment has contributed to energy efficiency in utilities and mining and to a shift away from energy-intensive activities in manufacturing and transportation industries. Technological advances have been most effective in increasing energy efficiency in construction and utilities and in decreasing energy-intensive activities in manufacturing industries. The results indicate that although efficiency contributes to a reduction in energy intensity in Canada, increasing activity in energy-intensive industries, such as oil and mining, partially offsets the efficiency gains in other industries.
Effect of cyclosporin A on interleukin-15-activated umbilical cord blood natural killer cell function.

BACKGROUND Interleukin (IL)-15-activated natural killer (NK) cells may provide a graft-versus-leukemia (GvL) effect post-umbilical cord blood (CB) transplantation. The effect of cyclosporin A (CsA), a calcineurin-inhibitor used for prophylaxis of graft-versus-host disease (GvHD), on IL-15-mediated activation, cytotoxic function and target-induced apoptosis of CB NK cells, was examined in comparison with adult peripheral blood (APB) NK cells. METHODS CsA was added to anti-CD3+/-IL-15-stimulated CB and APB mononuclear cells (MNC) for a 5-day incubation. CD3- CD56+ NK cell recovery was determined by flow cytometric analysis. Magnetic bead-purified CB and APB NK cells were stimulated with IL-15 for 18 h under the influence of CsA. NK activation (CD69), K562 cytotoxicity and NK-K562 interactions (CD54, perforin and annexin-V expression 4 h following contact with K562 cells) were assessed by flow cytometry. RESULTS CsA decreased CD3- CD56+ NK cell recovery in anti-CD3-stimulated CB MNC 5-day cultures, an effect that could be counteracted by IL-15; comparable effects were observed with APB. Short-term (18-h) experiments revealed that CsA down-regulated K562 cytotoxicity of IL-15-activated (P=0.018) but not resting (P=0.268) purified CB NK cells. IL-15-induced CB NK CD69 expression showed increased CsA sensitivity over APB (P=0.012). CsA down-regulated K562 cell-induced CD54 (P=0.028) but not perforin (P=0.416) expression of IL-15-activated CB NK cells. Target-induced apoptosis of IL-15-activated CB (P=0.043) but not APB (P=0.144) NK cells was decreased by CsA. DISCUSSION We have demonstrated differential CsA sensitivity of IL-15-activated CB and APB NK cells. These results may be used to improve the design of IL-15-activated NK cell adoptive immunotherapy in cancer patients receiving CsA post-CB transplantation.
Indirect selection of bacterial plasmids lacking identifiable phenotypic properties

A procedure is described that uses an indicator plasmid (pSC201) to identify cells in a bacterial population that have been co-transformed with a second plasmid lacking detectable phenotypic properties. Under appropriate conditions of indirect selection, between 50 and 85% of transformants carrying the indicator plasmid also contain the nonselected plasmid. A temperature-sensitive mutation in the replication functions of the indicator plasmid enables its elimination from doubly transformed bacteria. Using this procedure, we have isolated bacteria that carry only the small cryptic plasmid. P15A, of the Escherichia coli strain 15. This genetic element, which contains only 2,300 nucleotide pairs, is thus capable of functioning as a replicon independently of the two larger plasmids normally associated with it in E. coli 15 strains (Ikeda, Inuzuka, and Tomizawa, 1970).
Evaluation of selected endocrine disorders after anticancer treatment of solid tumors in childhood.

Wstęp. Coraz skuteczniejsze metody leczenia nowotworów wieku dziecięcego wpływają na lepszą przeżywalność, ale są przyczyną powstawania odległych następstw leczenia. Celem pracy jest ocena częstości występowania wybranych zaburzeń endokrynologicznych. Materiały i metody. Grupę badaną stanowiło 74 pacjentów (48 chłopców), w wieku 3,25-27 lat (średnia 13.24±6.21), co najmniej rok po zakończeniu leczenia przeciwnowotworowego z powodu guzów litych. Oceniono funkcję tarczycy, stężenie gonadotropin, lipidogram (cholesterol/LDL/TG, HDL) oraz BMD (z-score). Oceniono również parametry antropometryczne: wysokość SDS, masę ciała SDS oraz BMI SDS. Wyniki. Zaburzenia endokrynologiczne występowały u 66 (89,19%) pacjentów. U większości pacjentów (66,57%) obserwowano jedno lub dwa zaburzenia. Najczęstszymi nieprawidłowościami były dyslipidemia, występująca u 39 (58,21%) pacjentów, oraz nadwaga lub otyłość obserwowane u 25 (33,78%). Częstość występowania innych zaburzeń kształtowała się następująco: zaburzenia w stężeniu hormonów płciowych występowały u 14 (27,45%) pacjentów, nieprawidłowy poziom PTH u 13 (19,4%), nieprawidłowe stężenie IGF-1 u 9 (14,75%), niedobór wysokości u 9 (12,16%), niedobór masy ciała u 8 (11,11%), niedoczynność tarczycy u 7 (9,72%), obniżone BMD u 6 (8,69%), nadczynność tarczycy u 2 (2,78%). Wnioski. Pacjenci po leczeniu przeciwnowotworowym są w istotnym stopniu narażeni na zaburzenia endokrynologiczne. Wymagają oni szczegółowych badań kontrolnych w celu wykrycia nieprawidłowości oraz wdrożenia odpowiedniego leczenia. Warto zwrócić uwagę na częste występowanie więcej niż jednego zaburzenia u danego pacjenta.
Glycine N-Methyltransferase Deficiency Affects Niemann-Pick Type C2 Protein Stability and Regulates Hepatic Cholesterol Homeostasis

Nonalcoholic fatty liver disease (NAFLD) is associated with the development of metabolic syndromes and hepatocellular carcinoma (HCC). Cholesterol accumulation is related to NAFLD, whereas its detailed mechanism is not fully understood. Previously, we reported that glycine N-methyltransferase (GNMT) knockout (Gnmt-/-) mice develop chronic hepatitis and HCC. In this study, we showed that Gnmt-/- mice had hyperlipidemia and steatohepatitis. Single photon emission computed tomography images of mice injected with 131I-labeled 6β-iodocholesterol demonstrated that Gnmt-/- mice had slower hepatic cholesterol uptake and excretion rates than wild-type mice. In addition, genes related to cholesterol uptake (scavenger receptor class B type 1 [SR-B1] and ATP-binding cassette A1 (ABCA1)), intracellular trafficking (Niemann-Pick type C1 protein [NPC1] and Niemann-Pick type C2 protein [NPC2]) and excretion (ATP-binding cassette G1 [ABCG1]) were downregulated in Gnmt-/- mice. Yeast two-hybrid screenings and coimmunoprecipitation assays elucidated that the C conserved region (81–105 amino acids) of NPC2 interacts with the carboxyl-terminal fragment (171–295 amino acids) of GNMT. Confocal microscopy demonstrated that when cells were treated with low-density lipoprotein, NPC2 was released from lysosomes and interacts with GNMT in the cytosol. Overexpression of GNMT doubled the half-lives of both NPC2 isoforms and reduced cholesterol accumulation in cells. Furthermore, GNMT was downregulated in the liver tissues from patients suffering with NAFLD as well as from mice fed a high-fat diet, high-cholesterol diet or methionine/choline-deficient diet. In conclusion, our study demonstrated that GNMT regulates the homeostasis of cholesterol metabolism, and hepatic cholesterol accumulation may result from downregulation of GNMT and instability of its interactive protein NPC2. Novel therapeutics for steatohepatitis and HCC may be developed by using this concept
Challenges in Periprosthetic Knee-Joint Infection

The number of knee arthroplasty procedures is growing and projected to further increase. The risk for periprosthetic joint infection (PJI) is estimated to be low (<1%). However, considering the increasing number of total knee arthroplasty, the increasing number of patients with multiple comorbidities, and the lifelong risk for acquiring hematogenous infection, the total number of PJI will further increase. Despite existing treatment concepts for PJI of the knee, there are still questions to solve, such as type of debridement surgery in case of implant retention, the role of a spacer from a microbiological perspective, and the optimal duration of antimicrobial therapy. In this review, these questions will be analyzed according to the available literature and the experience of the authors. Moreover, we review the most recent data on infection, risk factors, and microbiology of PJI.
Managing Disruptions in Complex Adaptive Systems A Case for Resilience in Airline Operations Control

Introduction: Studies in naturalistic decision-making and resilience engineering have made significant advances towards characterising emergent cognitive behaviours in highly dynamic and distributed work settings. This doctoral project draws on such advances to examine disruptionmanagement processes that involve multiple players spanning multiple functions, expertises and differing levels of authority in the way each player influence the overall outcome. Method: This research integrates ethnographic and grounded theory methods in examining distributed decisionmaking, negotiations and coordination processes in airline operations control. Results and discussion: Preliminary findings suggest that complex networks of interdependencies—relating to resources and performance variables—across units involved in airline operations control promote reciprocity and the adoption of cooperative strategies. The findings further suggest that such emergent cognitive behaviours must be examined in context of the specific work setting studied. Such insights provide a framework for delineating how decision-makers collectively adapt formalised procedures in-flight while managing many-to-many mappings across conflicting goals.