Involvement of JAK2 and MAPK on type II nitric oxide synthase expression in skin-derived dendritic cells

In this report, we demonstrate that a fetal mouse skin-derived dendritic cell line produces nitric oxide (NO) in response to the endotoxin [lipopolysaccharide (LPS)] and to cytokines [tumor necrosis factor-alpha (TNF-alpha) and granulocyte-macrophage colony-stimulating factor (GM-CSF)]. Expression of the inducible isoform of NO synthase (iNOS) was confirmed by immunofluorescence with an antibody against iNOS. The tyrosine kinase inhibitor genistein decreased LPS- and GM-CSF-induced nitrite (NO(-2)) production. The effect of LPS and cytokines on NO(-2) production was inhibited by the Janus kinase 2 (JAK2) inhibitor tyrphostin B42. The p38 mitogen-activated protein kinase (p38 MAPK) inhibitor SB-203580 also reduced the NO(-2) production evoked by LPS, TNF-alpha, or GM-CSF, but it was not as effective as tyrphostin B42. Inhibition of MAPK kinase with PD-098059 also slightly reduced the effect of TNF-alpha or GM-CSF on NO(-2) production. Immunocytochemistry studies revealed that the transcription factor nuclear factor-kappaB was translocated from the cytoplasm into the nuclei of fetal skin-derived dendritic cells (FSDC) stimulated with LPS, and this translocation was inhibited by tyrphostin B42. Our results show that JAK2 plays a major role in the induction of iNOS in FSDC

Differential activation of nuclear factor kappa B subunits in a skin dendritic cell line in response to the strong sensitizer 2,4-dinitrofluorobenzene

Dendritic cell (DC) maturation is essential for the initiation of T-dependent immune responses. Nuclear factor kappa B (NF-kappaB) transcription factors are ubiquitously expressed signalling molecules, known to regulate the transcription of a large number of genes involved in immune responses, including cytokines and cell surface molecules. In this work, we studied the time-dependent activation of five members of the NF-kappaB family, p50, p52, p65, RelB and cRel, in a mouse skin DC line in response to stimulation with the strong sensitizer, 2,4-dinitrofluorobenzene (DNFB). Western blot assay revealed that exposure of fetal skin DC (FSDC) to DNFB induced the degradation of the inhibitor of NF-kappaB (IkappaB). Three out of its five members, i.e. p50, p52, and RelB, were similarly activated upon DNFB stimulation, with subsequent translocation of these subunits from the cytosol to the nucleus, but with different kinetics. In contrast, p65 expression was diminished in both the nucleus and the cytosol. The electrophoretic mobility shift assay (EMSA) showed that exposure of FSDC to DNFB induced DNA binding to NF-kappaB. Together, these results show that DNFB differentially activates the various members of the NF-kappaB family in skin DC

Granulocyte-macrophage colony-stimulating factor activates the transcription of nuclear factor kappa B and induces the expression of nitric oxide synthase in a skin dendritic cell line.

Nitric oxide (NO) produced by skin dendritic cells and keratinocytes plays an important role in skin physiology, growth and remodelling. Nitric oxide is also involved in skin inflammatory processes and in modulating antigen presentation (either enhancing or suppressing it). In this study, we found that GM-CSF stimulates the expression of the inducible isoform of nitric oxide synthase (iNOS) in a fetal-skin-derived dendritic cell line (FSDC) and, consequently, increases the nitrite production from 11.9 +/- 3.2 micromol/L (basal level) to 26.9 +/- 4.2 micromol/L. Pyrrolidinedithiocarbamate (PDTC) inhibits nitrite production, with a half maximal inhibitory concentration (IC50) of 19.3 micromol/L and the iNOS protein expression in FSDC. In addition, western blot assays revealed that exposure of FSDC to GM-CSF induces the phosphorylation and degradation of the inhibitor of NF-kappaB (IkB), with subsequent translocation of the p50, p52 and RelB subunits of the transcription nuclear factor kappa B (NF-kappaB) from the cytosol to the nucleus. Electrophoretic mobility shift assays (EMSA) showed that FSDC exposure to GM-CSF activates the transcription factor NF-kappaB. Together, these results show that GM-CSF induces iNOS expression in skin dendritic cells by a mechanism involving activation of the NF-kappaB pathway

Disclosing 3' UTR cis-elements and putative partners involved in gene expression regulation in Leishmania spp.

To identify putative cis-elements involved in gene expression regulation in Leishmania, we previously conducted an in silico investigation to find conserved intercoding sequences (CICS) in the genomes of L. major, L. infantum, and L. braziliensis. Here, the CICS databank was explored to search for sequences that were present in the untranslated regions (UTRs) of groups of genes showing similar expression profiles during in vitro differentiation. Using a selectable marker as a reporter gene, flanked by either an intact 3' UTR or a UTR lacking the conserved element, the regulatory role of a CICS was confirmed. We observed that the pattern of modulation of the mRNA levels was altered in the absence of the CICS. We also identified putative CICS RNA-binding proteins. This study suggests that the publicly available CICS database is a useful tool for identifying regulatory cis-elements for Leishmania genes and suggests the existence of post-transcriptional regulons in Leishmania

Variations in Inflammatory Markers in Acute Myocardial Infarction: a Longitudinal Study

Actualmente a inflamação é considerada uma componente importante na aterosclerose, desde o seu início até à ruptura da placa seguida de trombose e da progressiva obstrução do vaso. A ruptura da cápsula fibrótica da placa expõe factores de tecido presentes no seu núcleo necrótico que induzem o processo inflamatório, promovendo a adesão celular e a coagulação e que conduzem à formação do trombo. Por seu turno, várias citocinas e moléculas de adesão celular contribuem activamente para o desenvolvimento da placa. Em particular a citocina TNF-a e a molécula de adesão intercelular (ICAM-1) poderão ser indicadoras de inflamação enquanto que as formas solúveis de P-selectina e de CD40 ligando (sCD40L) poderão dar a magnitude da activação plaquetária. Neste trabalho foram estudados 17 doentes com enfarte de miocárdio submetidos a angioplastia (grupo AMI) e 16 doentes com confirmação angiográfica de ausência de doença coronária. Os doentes do grupo AMI foram seguidos nas primeiras 24h de evolução do enfarte agudo de miocárdio antes da administração de medicação e da intervenção angiográfica e ao longo do período de recuperação, 2 e 40 dias após enfarte. Foram medidas no soro por imunoensaio as concentrações de TNF-a e das formas solúveis de CD40L, ICAM-1 e P-selectina. Foram observadas variações significativas de sP-selectina relativamente aos controlos. Imediatamente após o enfarte de miocárdio verificou-se um aumento de sP-selectina, seguido de uma descida brusca dos seus níveis às 48h, e de um incremento para valores idênticos aos observados no grupo de controlo ao 40º dia. As variações observadas nas concentrações de sCD40L não foram significativas relativamente aos controlos. No entanto, verificou-se uma tendência de diminuição da concentração até 48h após o enfarte de miocárdio, seguindo-se um aumento que atingiu valores ligeiramente superiores ao do grupo controlo no 40º dia. As concentrações de TNF-a medidas foram sistematicamente superiores às verificadas no grupo controlo, tendo-se ainda observado uma subida gradual desde o enfarte de miocárdio até ao 40º dia, sendo este incremento significativo. Os valores de sICAM-1 não apresentaram quaisquer variações após o enfarte nem relativamente ao grupo controlo. As variações observadas sugerem um papel importante destes marcadores no processo inflamatório e na evolução do enfarte de miocárdio. O aumento brusco da concentração de sP-selectina após o enfarte de miocárdio evidencia a activação plaquetária e trombose. Na evolução do enfarte, e à medida que as variáveis hemodinâmicas retornam a valores estáveis, devido à medicação aplicada, o aumento de sCD40L e TNF-a em circulação pode reflectir o papel destas moléculas na recuperação endotelial e do miocárdio

Analysis of drugs returned by inpatient services after unit dose distribution in a portuguese public hospital

Unit-dose has been considered the most effective dispensing system in hospital pharmacy, however not all drugs are administered, are then returned to the pharmacy. The analysis of non-administered drugs might provide important data regarding pharmacotherapeutic follow-up, but also regarding pharmacy management decisions. The present study aims at depicting the drugs returned to the pharmacy following their previous unit-dose dispensing. Methods: During a period of 45 days, the unused returned drugs of five different inpatient clinical services were analyzed regarding the state of conservation, justification for return, inpatient clinical service provenance, and dosage regimen. Of a total of 65280 unit-dose dispensed drugs, 25.2% were returned (n=16431) and 74.9% of SOS (i.e. medications prescribed as needed) drugs (n=6583) were unused. Excluding SOS drugs, more than a half of the returned drugs (52.4%, n=4967), were probably returned due to unintended omission of administration, after excluding patients that were not physically on the unit and patients whose treatments were modified. The large majority of returned drugs (98.6%, n=16201) were suitable for reintroduction in the medication circuit. In order to accomplish the basic principles of unit-dose dispensing genesis, the returned drugs must be kept to a minimum. Therefore, the suspension of dispensing SOS drugs by unit-dose should be considered. Additionally, the careful analysis of returned drugs should be promoted, in order to avoid, as much as possible, the omission of administration.info:eu-repo/semantics/publishedVersio

Estilos de vida dos estudantes no ensino superior – consumo de tabaco.

info:eu-repo/semantics/publishedVersio

InSync

Version 1.0 ====================================== Abstract: ====================================== The InSync data set was collected at the Pervasive Computing lab at Ulster University. It consists of subjects performing activities of daily living (ADLs) in an atmosphere that mimics a real-life environment while data is collected using three different sensing technologies: inertial, image, and audio. The data set can be used to research human activity recognition algorithms to tackle problems on classification, transfer learning, data fusion, data segmentation, feature extraction, so on and so forth. Number of instances: ====================================== 16,959 (inertial data points) + 650 (thermal images) + 16,986 (audio files) Relevant information: ====================================== InSync contains 12 hours of data from ten subjects, consisting of 78 runs (times that a subject performed the scripted protocol). Sensor data from three different technologies (inertial, images and audio) captured the performance (not simulation) of the subjects performing ADLs. All the activities were annotated a posteriori using a video stream. ***** ACTIVITIES OF DAILY LIVING As the data set aimed at recording the subject's physical activity performance. The tasks consisted of ADLs and well-known scenarios. Three general scenarios were chosen, a bedroom-related scenario in which the subjects performed two of the ADLs, namely, personal hygiene and dressing, a breakfast-related scenario was chosen to embrace the ADL of feeding as it has extensively been used in literature, and free of obstacle scenario in which the subjects can walk alongside to demonstrate their transferring capabilities. The script was designed with nine high-level activities: Bedroom: (1) Napping (2) Wearing joggers (3) Combing hair (4) Brushing teeth Corridor: (5) Operating door Kitchen: (6) Drinking water (7) Eating cereal Livingroom: (8) Transporting (i.e. walking) (9) Resting (i.e. sitting in a chair) Details of the room's dimensions and sensor locations are available in the Relevant Papers. ***** SENSING TECHNOLOGY The deployed sensing technology included thirteen shimmer devices enabled with 3-axis accelerometers, four Matrix Voice ESP32 consisting of eight embedded microphones and four Thermal Vision Sensor (TVS). The sensing technology was placed as described next: Shimmers wore by the subject: - Right wrist - Left wrist - Lower back - Upper back - Right shoe Shimmers mounted on everyday items: - Comb - Toothbrush - Glass - Spoon - Jogger - Belt - Strap to mimic a watch - Strap to mimic smart shoe Matrix Voice ESP32 (one located in each room): - Bedroom - Corridor - Kitchen - Livingroom Thermal sensor (one located in each room): - Bedroom - Corridor - Kitchen - Livingroom Attribute information: ====================================== The data set comprises the readings of inertial sensors, thermal images, and audio files to recorded performed ADLs. There is a total of 60 attributes for the inertial data which includes the mean value and root-mean-square (RMS) from x, y, and z-axis. The thermal data consists of grayscale images in 32x32 pixels, and the audio data consists of 44.1 kHz waveform audio files. A list of videos of the experiment can be seen in the following links. Bedroom: (1) napping: https://youtu.be/IqWLKsgch6A (2) wearing joggers: https://youtu.be/FJBjO9C4Q4U (3) combing hair: https://youtu.be/bYKrKbVBNos (4) brushing teeth: https://youtu.be/wuVkrWlsmSs Corridor: (5) operating door: https://youtu.be/pWJjx3TH6Q4 Kitchen: (6) drinking water: https://youtu.be/wS9OBKK_LFY (7) eating cereal: https://youtu.be/nOK8TuyCXBA Livingroom: (8) transporting (i.e. walking): https://youtu.be/45MGsYS9cYg (9) resting (i.e. sitting in a chair): https://youtu.be/45MGsYS9cYg IMPORTANT: The videos previously provided were recorded using conventional webcams. The videos were used as ground truth; they were not used for training nor testing purposes. Note that the participant's identity has been considered by blurring their face. The speed of the videos varies as different sampling rates were used when recording the videos