Miracle-dz/UniFAR-FTUs huggingface dataset:
Mutants of Interleukin 13 with Altered Reactivity toward Interleukin 13 Receptors*

Interleukin 13 (IL13) belongs to a family of cytokines whose members exhibit structural homology, despite amino acid sequence dissimilarity. For example, while of limited sequence homology, IL13 and IL4 share a signaling receptor, IL13/4 receptor, on a variety of human normal cells. However, a subclass of IL4-independent IL13 receptors is overexpressed on certain transformed cells, including human malignant gliomas. We introduced mutations into human (h) IL13 to determine the site(s) involved in interaction with the shared receptor and/or the glioma-associated receptor. This analysis identified at least three protein regions that are needed for signaling through the shared receptor. These regions were localized to α-helices A, C, and D and were mainly separate from the region(s) needed to interact with the glioma-associated receptor. Glutamic acids at positions 13 and 16 in hIL13 α-helix A, arginine and serine at positions 66 and 69 in helix C, and arginine at position 109 in helix D were found to be important in inducing biological signaling since their specific mutation resulted in loss and/or gain of function phenomena. We demonstrate that the molecular requirements of hIL13 to interact with its respective receptors are generally distinct and can be controlled by mutagenesis of the cytokine.
Protein Kinase C-mediated Phosphorylation and Functional Regulation of Dopamine Transporters in Striatal Synaptosomes*

Dopamine transporters (DATs) are members of a family of Na+- and Cl−-dependent neurotransmitter transporters responsible for the rapid clearance of dopamine from synaptic clefts. The predicted primary sequence of DAT contains numerous consensus phosphorylation sites. In this report we demonstrate that DATs undergo endogenous phosphorylation in striatal synaptosomes that is regulated by activators of protein kinase C. Rat striatal synaptosomes were metabolically labeled with [32P]orthophosphate, and solubilized homogenates were subjected to immunoprecipitation with an antiserum specific for DAT. Basal phosphorylation occurred in the absence of exogenous treatments, and the phosphorylation level was rapidly increased when synaptosomes were treated with the phosphatase inhibitors okadaic acid or calyculin. Treatment of synaptosomes with the protein kinase C activator phorbol 12-myristate 13-acetate (PMA) also increased the level of phosphate incorporation. This occurred within 10 min and was dosedependent between 0.1 and 1 μm PMA. DAT phosphorylation was also significantly increased by two other protein kinase C activators, (−)-indolactam V and 1-oleoyl-2-acetyl-sn-glycerol. The inactive phorbol ester 4α-phorbol 12,13-didecanoate at 10 μm was without effect, and PMA-induced phosphorylation was blocked by treatment of synaptosomes with the protein kinase C inhibitors staurosporine and bisindoylmaleimide. These results indicate that DATs undergo rapidin vivo phosphorylation in response to protein kinase C activation and that a robust mechanism exists in synaptosomes for DAT dephosphorylation. Dopamine transport activity in synaptosomes was reduced by all treatments that promoted DAT phosphorylation, with comparable dose, time, and inhibitor characteristics. The change in transport activity was produced by a reduction inV max with no significant effect on theK m for dopamine. These results suggest that synaptosomal dopamine transport activity is regulated by phosphorylation of DAT and present a potential mechanism for local neuronal control of synaptic neurotransmitter levels and consequent downstream neural activity.
Graphene oxide as an anaerobic membrane scaffold for the enhancement of B. adolescentis proliferation and antagonistic effects against pathogens E. coli and S. aureus

The impact of the gut microbiota on human health is widely perceived as the most exciting advancement in biomedicine. The gut microbiota has been known to play a crucial role in defining states of human health and diseases, and thus becomes a potential new territory for drug targeting. Herein, graphene oxide (GO) interaction with five common human gut bacteria, B. adolescentis, L. acidophilus, E. coli, E. faecalis, and S. aureus, was studied. It was shown that, in bacterial media, GO sheets were able to form effective, anaerobic membrane scaffolds that enhanced the antagonistic activity of B. adolescentis against the pathogens E. coli andS. aureus. Data obtained using bacterial growth measurements, colony counting and 16S rRNA gene sequencing consistently indicated that GO sheets promoted proliferation of gut bacteria, particularly for B. adolescentis. Scanning electron microscopy, atomic force microscopy images, and membrane potential measurements showed that cell membranes maintained their integrity and that no observable variations in cell morphology were induced after interaction with GO sheets, indicating good biocompatibility of GO. These results suggest the possibility of using GO sheets as efficient drug carriers in therapeutic applications to treat diseases related to the gut microbiota.
Capacity after stroke.

The new knowledge and clearer concepts of the pathogenesis of cerebrovascular disease and of "order and disorder" in the cerebral circulation acquired in recent years have improved the possibilities of treatment, which may now be considered at four stages in the evolution of a stroke: by prevention, when factors contributing to "stroke-proneness" may be identified and offset; by anticipation of impending stroke and the use of new techniques to locate and treat disordered regional blood flow in the cerebral circulation; by medical or surgical modi fication of progressing stroke; and by rehabilitation of completed stroke. There are two phases of the latter: the first a period of intensive care required at onset to save life and ensure survival: the second, often prolonged, being the treat ment required to obtain the best recovery possible for a disabled survivor.
An in situ study of the nanomechanical properties of barnacle (Balanus amphitrite) cyprid cement using atomic force microscopy (AFM)

Abstract Cyprids are the final planktonic stage in the larval dispersal of barnacles and are responsible for surface exploration and attachment to appropriate substrata. The nanomechanical properties of barnacle (Balanus amphitrite) cyprid permanent cement were studied in situ using atomic force microscopy (AFM). Force curves were recorded from the cement disc continually over the course of its curing and these were subsequently analysed using custom software. Results showed a narrowing of the pull-off force distribution with time, as well as a reduction in molecular stretch length over time. In addition, there was a strong correlation between maximum pull-off force and molecular stretch length for the cement, suggesting ‘curing’ of the adhesive; some force curves also contained a ‘fingerprint’ of modular protein unfolding. This study provides the first direct experimental evidence in support of a putative ‘tanning’ mechanism in barnacle cyprid cement.
Protective effects of propofol on acute lung injury induced by oleic acid in conscious rats*

Objectives:Oleic acid has been used to induce acute lung injury (ALI) in animals. In patients with acute respiratory distress syndrome (ARDS), the blood level of oleic acid was increased. The mechanism and therapeutic regimen of ARDS and oleic acid-induced ALI remain undefined. In the present study, we investigated the oleic acid-induced changes in lung variables for the measure of ALI, inflammatory mediators, and neutrophil-derived substances. We evaluated the effects of pretreatment and posttreatment with propofol. Design:Randomized, controlled animal study. Setting:University research laboratory. Subjects:Fifty adult male Sprague-Dawley rats weighing 250–300 g. Interventions:We employed a conscious and unrestrained rat model. Oleic acid at a dose of 100 mg/kg was administered intravenously. Propofol (30 mg/kg) was given by intravenous infusion (6 mg/kg/min for 5 mins) 30 mins before (pretreatment) and 30 mins after (posttreatment) oleic acid. Measurements and Main Results:We monitored the arterial pressure, heart rate, and blood gas. The lung weight changes, exhaled nitric oxide, protein concentration in bronchoalveolar lavage, and Evans blue content in lung tissue were determined. The plasma nitrate/nitrite, methylguanidine, cytokines (tumor necrosis factor-α, interleukin-1β, interleukin-6, and interleukin-10), neutrophil elastase, myeloperoxidase, malondialdehyde, and sodium- and potassium-activated adenosine triphosphatase (Na+-K+-ATPase) were detected. Histopathological examination of the lung was performed. Oleic acid caused systemic hypotension and severe ALI as evidenced by the increases in the extent of ALI, impairment of pulmonary functions (blood gas variables), and lung pathology. In addition, oleic acid significantly increased inflammatory mediators and neutrophil-derived factors but depressed Na+-K+-ATPase. The inducible nitric oxide synthase was up-regulated. Pre- or posttreatment with propofol was capable of reversing the oleic acid-induced changes and attenuating the extent of ALI. Conclusions:Oleic acid resulted in sepsis-like responses including ALI, inflammatory reaction, and increased neutrophil-derived factors. It depressed the Na+-K+-ATPase activity but up-regulated inducible nitric oxide synthase. Treatment with propofol abrogated or reversed the oleic acid-induced changes.
Incidence of knee sepsis after ACL reconstruction at one institution: the impact of a clinical pathway.

BACKGROUND After experiencing an unusually high incidence of knee sepsis after anterior cruciate ligament (ACL) reconstruction, we sought to (1) describe how we resolved this problem through temporary discontinuation of the procedure, formation of a multidisciplinary ACL Task Force, systematic investigation of clinical data and institutional care practices, and development and implementation of an evidence-based ACL Clinical Pathway (the Pathway); and (2) report our findings and results. METHODS From 1999 through 2008, thirty-seven cases of knee sepsis after ACL reconstruction were recorded at our institution. In 2008 (yearly incidence, 4.4%), ACL reconstructions were temporarily suspended and a Task Force was assembled to (1) identify infection risk factors or epidemiological links among cases, (2) inspect environment and processes for possible infection sources, and (3) update existing perioperative practices according to current evidence-based guidelines to reduce surgical site infection risk. These actions led to the development of the Pathway for patients and providers. The rates of knee sepsis before and after the Pathway was implemented were compared. RESULTS There was no consistent risk factor or epidemiologic link among the cases of knee sepsis other than the time and place of the ACL reconstruction. Process review identified shortfalls in decontamination and sterilization of some surgical equipment. Perioperative care practices review revealed wide interprovider variation. Pathway implementation reduced the rate of knee sepsis after ACL reconstruction from 1.96% (twenty-four cases after 1226 ACL reconstructions performed from 2002 to 2008) to 0% (zero cases after 500 ACL reconstructions performed from 2008 to 2011); the difference was significant (p = 0.003). CONCLUSIONS When a Task Force investigation suggested that knee sepsis after ACL reconstruction was a multifactorial problem, we implemented and standardized evidence-based perioperative care practices via the institution-wide Pathway, which significantly improved the quality and consistency of care for patients undergoing ACL reconstruction, as well evidenced by the elimination of knee sepsis.
Efficacy versus Effectiveness

Archie Cochrane, British famous clinical epidemiologist, defined two concepts related to assessing healthcare interventions. Efficacy is the extent to which an intervention does more good than harm under ideal circumstances. Effectiveness assesses whether an intervention does more good than harm when provided under usual circumstances of healthcare practice.1) Research on efficacy and effectiveness has also been described as explanatory and pragmatic trials. When defining the research question, it must be considered whether one asking a pragmatic or an explanatory question. This will have important implications for study design.2) Pragmatic research assesses whether an intervention works under real-life conditions and whether it works in terms that matter to the patient. Explanatory research assesses whether an intervention works under ideal or selected conditions. The distinction between pragmatic and explanatory research is important because it will determine key methodological issues relating to patient selection, definition of the intervention and controls, use of blinding and placebos, choice of outcome measure, and type of analysis.3) Efficacy studies are frequently conducted in large tertiary care, referral settings, which tend to have more specialized clinicians and better technical equipment than primary care facilities. For effectiveness trials, settings should reflect the initial care facilities available to a diverse population with the condition of interest. Therefore, populations in primary care may be an adequate subject group for effectiveness studies.4) The Korean Journal of Family Medicine (KJFM) publishes research and reviews related to primary care, and those related to clinical and epidemiological health services. This means that the KJFM is focused primarily on the publication of effectiveness research. The distinctions between pragmatic and explanatory research pertain to patient selection, definition of the intervention and controls, and choice of outcome measure. Effectiveness research has several characteristics: 1) patient selection should reflect routine practice; 2) the intervention should be provided by normal staff with routine training; 3) the control may therefore be routine practice rather than a placebo; 4) outcome measures are usually patient centered and incorporate broad measures of health-related quality of life.5) The editorial board, reviewer, and authors must all have such an understanding to achieve the aims and scope of the KJFM.
A Comparison of Axonal and Somatodendritic Dopamine Release Using In Vivo Dialysis

Abstract: The release of endogenous dopamine from the axon terminal field in the nucleus accumbens and from the A10 dopamine cell bodies of conscious rats was measured using intracranial dialysis. Release of dopamine from both areas was calcium‐dependent and markedly inhibited by the presence of the D2 agonist, quinpirole, in the dialysis buffer. However, the addition of tetrodotoxin to the buffer produced less of a decrease in dopamine in the A10 region than in the nucleus accumbens. When dopamine release was examined by substituting K+ for Na+ or by adding amphetamine to the buffer, the evoked release was significantly less in the A10 region than in the nucleus accumbens. Likewise, enhanced extracellular dopamine following blockade of reuptake by nomifensine addition to the dialysis buffer was not as great in the A10 region as in the nucleus accumbens. These data argue that, in general, axonal and somatodendritic dopamine release are regulated by similar factors, although somatodendritic release is less influenced by action potential generation and less responsive to some releasing agents.
Sorting of Mannose 6-Phosphate Receptors Mediated by the GGAs

The delivery of soluble hydrolases to lysosomes is mediated by the cation-independent and cation-dependent mannose 6-phosphate receptors. The cytosolic tails of both receptors contain acidic-cluster-dileucine signals that direct sorting from the trans-Golgi network to the endosomal-lysosomal system. We found that these signals bind to the VHS domain of the Golgi-localized, γ-ear–containing, ARF-binding proteins (GGAs). The receptors and the GGAs left the trans-Golgi network on the same tubulo-vesicular carriers. A dominant-negative GGA mutant blocked exit of the receptors from the trans-Golgi network. Thus, the GGAs appear to mediate sorting of the mannose 6-phosphate receptors at the trans-Golgi network.
dpr and sod in Streptococcus mutans Are Involved in Coexistence with S. sanguinis, and PerR Is Associated with Resistance to H2O2

ABSTRACT Large numbers of bacteria coexist in the oral cavity. Streptococcus sanguinis, one of the major bacteria in dental plaque, produces hydrogen peroxide (H2O2), which interferes with the growth of other bacteria. Streptococcus mutans, a cariogenic bacterium, can coexist with S. sanguinis in dental plaque, but to do so, it needs a means of detoxifying the H2O2 produced by S. sanguinis. In this study, we investigated the association of three oxidative stress factors, Dpr, superoxide dismutase (SOD), and AhpCF, with the resistance of S. sanguinis to H2O2. The knockout of dpr and sod significantly increased susceptibility to H2O2, while the knockout of ahpCF had no apparent effect on susceptibility. In particular, dpr inactivation resulted in hypersensitivity to H2O2. Next, we sought to identify the factor(s) involved in the regulation of these oxidative stress genes and found that PerR negatively regulated dpr expression. The knockout of perR caused increased dpr expression levels, resulting in low-level susceptibility to H2O2 compared with the wild type. Furthermore, we evaluated the roles of perR, dpr, and sod when S. mutans was cocultured with S. sanguinis. Culturing of the dpr or sod mutant with S. sanguinis showed a significant decrease in the S. mutans population ratio compared with the wild type, while the perR mutant increased the ratio. Our results suggest that dpr and sod in S. mutans are involved in coexistence with S. sanguinis, and PerR is associated with resistance to H2O2 in regulating the expression of Dpr.
Mycobacterium bovis Bacillus Calmette-Guérin Induces TLR2-Mediated Formation of Lipid Bodies: Intracellular Domains for Eicosanoid Synthesis In Vivo1

Differentiation of macrophages into foamy (lipid-laden) macrophages is a common pathological observation in tuberculous granulomas both in experimental settings as well as in clinical conditions; however, the mechanisms that regulate intracellular lipid accumulation in the course of mycobacterial infection and their significance to pathophysiology of tuberculosis are not well understood. In this study, we investigated the mechanisms of formation and function of lipid-laden macrophages in a murine model of tuberculosis. Mycobacterium bovis bacillus Calmette-Guérin (BCG), but not Mycobacterium smegmatis, induced a dose- and time-dependent increase in lipid body-inducible nonmembrane-bound cytoplasmic lipid domain size and numbers. Lipid body formation was drastically inhibited in TLR2-, but not in TLR4-deficient mice, indicating a role for TLR2 in BCG recognition and signaling to form lipid bodies. Increase in lipid bodies during infection correlated with increased generation of PGE2 and localization of cyclooxygenase-2 within lipid bodies. Moreover, we demonstrated by intracellular immunofluorescent localization of newly formed eicosanoid that lipid bodies were the predominant sites of PGE2 synthesis in activated macrophages. Our findings demonstrated that BCG-induced lipid body formation is TLR2 mediated and these structures function as signaling platforms in inflammatory mediator production, because compartmentalization of substrate and key enzymes within lipid bodies has impact on the capacity of activated leukocytes to generate increased amounts of eicosanoids during experimental infection by BCG.
Developmental Expression of the Myelin‐Associated Glycoprotein in the Peripheral Nervous System Is Different from That in the Central Nervous System

Abstract: We recently characterized two developmentally regulated myelin‐associated glycoprotein (MAG) polypeptides synthesized by mouse brain mRNA in vitro. We now extended these studies to include the peripheral nervous system (PNS). Total cytoplasmic RNA was isolated from the sciatic nerves of 7‐, 12‐, and 17‐day‐old and adult rats and translated in vitro in a rabbit reticulocyte lysate system. In contrast to results in the CNS, it appears that only one MAG polypeptide, p67MAG, is synthesized by PNS mRNA at all ages. The implications of these findings are discussed with respect to recent observations concerning both the localization of MAG and the synthesis of MAG in the PNS of dysmyelinating mutant mice.
Design of nonlinear control systems by means of differential genetic programming

Based on the genetic programming (GP), a new design method of optimal and/or robust controllers of nonlinear systems is proposed. First we introduce a new type of genetic programming, which is called "differential GP". The differential GP combines the conventional GP with an automatic differentiation scheme, so that it can solve Hamilton-Jacobi-Bellman (HJB)/Hamilton-Jacobi-Isaacs (HJI)/Francis - Byrnes - Isidori (FBI) equations. Our design method is mainly based on the differential GP technique, and so it gives us a new possibility of the HJB/HJI/FBI based design techniques. Lastly, the effectiveness of the differential GP based design method is demonstrated through some design examples of nonlinear systems.
Efectos de la crisis económica sobre el emprendimiento en empresas de economía social en España: un análisis espacial

En los ultimos anos, la desfavorable evolucion de la coyuntura economica ha provocado intensos cambios estructurales y espaciales en la creacion de empresas que cuestionan la validez de algunos supuestos basicos sobre dinamica emprendedora y desarrollo territorial. Por ello, el objetivo de este trabajo es analizar la existencia de patrones espaciales de creacion de empresas de economia social en Espana y estudiar su evolucion temporal durante dos etapas de intensos cambios: una de crecimiento (2002-2007) y otra de profunda crisis generalizada (2008-2013). A partir de la definicion de un indice de agrupacion de empresas de economia social aplicamos tecnicas estadisticas basadas en econometria espacial (indice de Moran y LISA). Para tal fin se tomaron datos longitudinales del numero de empresas creadas en Espana, durante el periodo 2002-2013, bajo las formulas de economia social y mercantil comparandose posteriormente a nivel autonomico. Su dinamica se analiza conjuntamente con el PIB regional como indicador del contexto economico. Los resultados mas generales apuntan a que el patron de expansion de la dinamica emprendedora en empresas de economia social es negativo y regionalmente heterogeneo, mostrando la existencia de algunas concentraciones espaciales, en lo que podrian denominarse Clusters de Empresas de Economia Social . A la vista de esos resultados y patrones se evidencia una erosion en la influencia de determinados factores de tipo contextual (tradicion, cultura y normativa) en esa actividad emprendedora. Nuestras contribuciones tienen implicaciones para la academia y para los decisores de politicas publicas de apoyo a la creacion de empresas y la economia social.