Expression of mRNA for phospholipase A(2), cyclooxygenases, and lipoxygenases in cultured human umbilical vascular endothelial and smooth muscle cells and in biopsies from umbilical arteries and veins

Arachidonic acid (AA) is released by phospholipase A(2) (PLA(2)) and then converted into vasoactive and inflammatory eicosanoids by cyclooxygenases (COX) and lipoxygenases (LOX). These eicosanoids are important paracrine regulators of vascular permeability, blood flow, local pro- and anticoagulant activity and they play a major role in the local inflammatory response. We have investigated the presence of mRNAs for PLA(2) and for isoforms of COX and LOX in both human endothelial cells (EC) and in human smooth muscle cells (SMC) in culture and in vascular biopsies of human umbilical veins (HUVB) and arteries (HUAB) by using the reversed transcription-polymerase chain reaction (RT-PCR) technique. Results show detectable levels of PLA(2) type IV (cPLA(2)) in cultured EC and SMC and in vascular wall biopsies from HUAB and HUVB. The cultured EC and SMC demonstrate higher levels of both COX-1 and COX-2 with PCR analyses than do vascular wall biopsies from HUAB and HUVB. This indicates a difference in the native expression of COX-1 and COX-2 in cultures of EC and SMC compared to that in biopsies from intact vessel walls. The EC and SMC in culture do not express mRNA for 5-LOX, that was, however, expressed in the vascular wall biopsies. This speaks in favour of a constitutive, i.e, in vivo expression of 5-LOX in SMC in the vascular wall of both umbilical vein and arteries. Thus results from in vitro studies of constitutive COX and LOX expression in EC and vascular SMC in culture cannot simply be extrapolated to represent in vivo conditions

Different induction mechanisms of mRNA for inducible nitric oxide synthase in rat smooth muscle cells in culture and in aortic strips

AbstractThe expression of mRNA for the inducible form of nitric oxide synthase, (iNOS), was studied in rat aortic smooth muscle cells, (SMCs) in cell culture and in strips of rat aorta by reverse transcriptase coupled to the polymerase chain reaction. iNOS mRNA expression was weak in cultured SMCs when exposed to either interferon-γ (IFNγ) or lipopolysaccharide (LPS), but the combination LPS + IFNγ enhanced the expression. In aortic strips LPS alone induced a pronounced expression, with no further increase by IFNγ. Cycloheximide potentiated the expression of iNOS mRNA in SMCs in culture stimulated with LPS + IFNγ but attenuated the response in aortic strips.The results indicate different cellular signaling pathways for the induction of iNOS mRNA by LPS and/or IFNγ, in cultured SMCs and in rat aortic strips

Effect of D-Cycloserine on the Effect of Concentrated Exposure and Response Prevention in Difficult-to-Treat Obsessive-Compulsive Disorder: A Randomized Clinical Trial

Importance Evidence is lacking for viable treatment options for patients with difficult-to-treat obsessive-compulsive disorder (OCD). It has been suggested that D-cycloserine (DCS) could potentiate the effect of exposure and response prevention (ERP) treatment, but the hypothesis has not been tested among patients with difficult-to-treat OCD. Objective To evaluate whether DCS potentiates the effect of concentrated ERP among patients with difficult-to-treat OCD. Design, Setting, and Participants The study was a randomized placebo-controlled triple-masked study with a 12-month follow-up. Participants were adult outpatients with difficult-to-treat OCD. A total of 220 potential participants were referred, of whom 36 did not meet inclusion criteria and 21 declined to participate. Patients had either relapsed after (n = 100) or not responded to (n = 63) previous ERP treatment. A total of 9 specialized OCD teams within the public health care system in Norway participated, giving national coverage. An expert team of therapists from the coordinating site delivered treatment. Inclusion of patients started in January 2016 and ended in August 2017. Data analysis was conducted February to September 2019. Interventions All patients received individual, concentrated ERP treatment delivered during 4 consecutive days in a group setting (the Bergen 4-day treatment format) combined with 100 mg DCS, 250 mg DCS, or placebo. Main outcomes and Measures Change in symptoms of OCD and change in diagnostic status. Secondary outcomes measures included self-reported symptoms of OCD, anxiety, depression, and quality of life. Results The total sample of 163 patients had a mean (SD) age of 34.5 (10.9) years, and most were women (117 [71.8%]). They had experienced OCD for a mean (SD) of 16.2 (10.2) years. A total of 65 patients (39.9%) were randomized to receive 100 mg DCS, 67 (41.1%) to 250 mg of DCS, and 31 (19.0%) to placebo. Overall, 91 (56.5%) achieved remission at posttreatment, while 70 (47.9%) did so at the 12-month follow-up. There was no significant difference in remission rates among groups. There was a significant reduction in symptoms at 12 months, and within-group effect sizes ranged from 3.01 (95% CI, 2.38-3.63) for the group receiving 250 mg DCS to 3.49 (95% CI, 2.78-4.18) for the group receiving 100 mg DCS (all P < .001). However, there was no significant effect of treatment group compared with placebo in obsessive-compulsive symptoms (250 mg group at posttreatment: d = 0.33; 95% CI, −0.10 to 0.76; 100 mg group at posttreatment: d = 0.36; 95% CI, −0.08 to 0.79), symptoms of depression and anxiety (eg, Patient Health Questionnaire–9 score among 250 mg group at 12-month follow-up: d = 0.30; 95% CI, −0.17 to 0.76; Generalized Anxiety Disorder–7 score among 100 mg group at 12-month follow-up: d = 0.27; 95% CI, −0.19 to 0.73), and well-being (250 mg group: d = 0.10; 95% CI, −0.42 to 0.63; 100 mg group: d = 0.34; 95% CI, −0.19 to 0.86). No serious adverse effects were reported. Conclusions and Relevance In this study, DCS did not potentiate ERP treatment effect, but concentrated ERP treatment was associated with improvement. Trial Registration ClinicalTrials.gov identifier: NCT02656342publishedVersio

The feasibility of an augment reality system to study the psychophysiological correlates of fear-related responses

Previous studies have successfully used augmented reality (AR) as an aid to exposure-based treatments for anxiety disorders. However, to the best of our knowledge, none of these studies have measured the physiological correlates of the fear response, relying solely on self-reports and behavioral avoidance tests.publishe

Histone Variants and Their Post-Translational Modifications in Primary Human Fat Cells

Epigenetic changes related to human disease cannot be fully addressed by studies of cells from cultures or from other mammals. We isolated human fat cells from subcutaneous abdominal fat tissue of female subjects and extracted histones from either purified nuclei or intact cells. Direct acid extraction of whole adipocytes was more efficient, yielding about 100 µg of protein with histone content of 60% –70% from 10 mL of fat cells. Differential proteolysis of the protein extracts by trypsin or ArgC-protease followed by nanoLC/MS/MS with alternating CID/ETD peptide sequencing identified 19 histone variants. Four variants were found at the protein level for the first time; particularly HIST2H4B was identified besides the only H4 isoform earlier known to be expressed in humans. Three of the found H2A potentially organize small nucleosomes in transcriptionally active chromatin, while two H2AFY variants inactivate X chromosome in female cells. HIST1H2BA and three of the identified H1 variants had earlier been described only as oocyte or testis specific histones. H2AFX and H2AFY revealed differential and variable N-terminal processing. Out of 78 histone modifications by acetylation/trimethylation, methylation, dimethylation, phosphorylation and ubiquitination, identified from six subjects, 68 were found for the first time. Only 23 of these modifications were detected in two or more subjects, while all the others were individual specific. The direct acid extraction of adipocytes allows for personal epigenetic analyses of human fat tissue, for profiling of histone modifications related to obesity, diabetes and metabolic syndrome, as well as for selection of individual medical treatments

The importance of the altricial – precocial spectrum for social complexity in mammals and birds:A review

Various types of long-term stable relationships that individuals uphold, including cooperation and competition between group members, define social complexity in vertebrates. Numerous life history, physiological and cognitive traits have been shown to affect, or to be affected by, such social relationships. As such, differences in developmental modes, i.e. the ‘altricial-precocial’ spectrum, may play an important role in understanding the interspecific variation in occurrence of social interactions, but to what extent this is the case is unclear because the role of the developmental mode has not been studied directly in across-species studies of sociality. In other words, although there are studies on the effects of developmental mode on brain size, on the effects of brain size on cognition, and on the effects of cognition on social complexity, there are no studies directly investigating the link between developmental mode and social complexity. This is surprising because developmental differences play a significant role in the evolution of, for example, brain size, which is in turn considered an essential building block with respect to social complexity. Here, we compiled an overview of studies on various aspects of the complexity of social systems in altricial and precocial mammals and birds. Although systematic studies are scarce and do not allow for a quantitative comparison, we show that several forms of social relationships and cognitive abilities occur in species along the entire developmental spectrum. Based on the existing evidence it seems that differences in developmental modes play a minor role in whether or not individuals or species are able to meet the cognitive capabilities and requirements for maintaining complex social relationships. Given the scarcity of comparative studies and potential subtle differences, however, we suggest that future studies should consider developmental differences to determine whether our finding is general or whether some of the vast variation in social complexity across species can be explained by developmental mode. This would allow a more detailed assessment of the relative importance of developmental mode in the evolution of vertebrate social systems

Prediction value of genetic and neuromarkers in blood and liquor in patients with severe traumatic brain injury

Background: Severe traumatic brain injury (sTBI) is the most common cause of mortality in young adults. sTBI induces variable brain damage, invisible in Computer Tomographic scans early post-trauma. Further, neurology is difficult to evaluate in sedated patients. Therefore, biochemical neuromarkers (BNMs) in blood or cerebrospinal fluid (CSF) may be valuable tools to both evaluate trauma and to prognosticate patient outcome. Aims: The aim of the thesis was to evaluate if concentrations of the BNMs; Glial Fibrillary Acid Protein (GFAP, CSF, study IV), Neurofilament light (NFL, CSF, study IV), Tau (CSF, study II), β-amyloid (1-42) and amyloid precursor-proteins (CSF & plasma, study I) were enhanced after a sTBI. Further, we investigated if these levels were correlated to outcome, neurology and patient ability of daily living 1-year post-trauma. Finally, we explored if patient-genotype, specifically Apolipoprotein E, (gene APOE), influenced 1-year outcome in sTBI-patients, (plasma, study III). Methods: Patients were consecutively included if; aged ≥7 years, < 9 in Glasgow Coma Scale, receiving an indwelling ventricular catheter allowing CSF sampling), were artificially ventilated and admitted to the Neurointensive care unit (NICU) within 48h post-trauma. NICU-care was performed according to a standardized protocol. CSF samples were collected on days 0-4, 6, 8 and once on days 11-18. Surviving patients were assessed at 1-year evaluating; 1) outcome by Glasgow Outcome Scale (GOS), 2. neurology and 3. activities of daily living. NFL, GFAP, Tau, β-amyloid (1-42) and amyloid precursor-proteins were all analyzed by ELISAmethods. APOE genotyping was performed by polymerase chain reaction & solidphase mini-sequencing. Results: During the inclusion period, patients (n=28-96) were included into studies IIV for CSF and /or blood sampling. Study I; β-amyloid (1-42) and amyloid precursor-proteins increased from day 0 until day 11 in the CSF, but not in plasma. In study II we found enhanced levels of CSF-Tau on days 2-3 correlated to mortality (GOS 1) at 1-year. In study III we found that patients with APOE allele 4 had worse outcome (GOS) at 1-year. Finally, in paper IV we found increased CSF levels of GFAP and NFL both correlating to outcome (GOS) at 1-year. Conclusions; In this thesis we have found in sTBI-patients that genetic and BNMs in the plasma and/or CSF correlate to outcome at 1 -year post-trauma. The result may be clinically applicable to prognosticate outcome and influence treatment paradigms in these patients

http://urn.kb.se/resolve?urn=urn:nbn:se:liu:diva-104290 The Concentration of b-Carotene in Human Adipocytes, but Not the Whole-Body Adipocyte Stores, Is Reduced in

We have examined the concentration of b-carotene in the fat of isolated abdominal subcutaneous adipocytes obtained from lean (BMI,23 kg/m 2), non-obese with higher BMI (23#BMI,28 kg/m 2), obese (BMI$28 kg/m 2), and from a group of obese subjects with type 2 diabetes. The concentration of b-carotene was 50 % lower in the adipocytes from the obese and obese/diabetic groups compared with the lean and non-obese groups. Interestingly, the total amount of b-carotene in the adipocyte stores of each subject was constant among all groups. Triacylglycerol constituted 9261 % (by weight) of the adipocyte lipids in the lean group and this was increased to 9962 % in the obese group with diabetes (p,0.05). The concentration of cholesteryl esters was in all cases,0.1 g per 100 g of total lipids, demonstrating that mature human adipocytes have negligible stores of cholesteryl ester. Our findings demonstrate that adipocyte concentrations of b-carotene are reduced in obese subjects. The lower concentrations in adipocytes from subjects with type 2 diabetes apparently reflect subjects obesity. Our finding that whole-body stores of b-carotene in adipocytes are constant raises new questions regarding what function it serves, as well as the mechanisms for maintaining constant levels in the face of varie

Innovativ kostnadsberäkningsmodell för additiv tillverkning : En kvalitativ fallstudie med bidrag till utvecklingen av självkostnadsberäkning

Actors within the manufacturing industry are constantly looking for innovative manufacturing methods to remain competitive. Additive manufacturing (AM), also referred to as 3D printing, is considered to have a good potential in the development towards industry 4.0. Though the cost control of AM-processes is an important step towards future business opportunities, little research has been implemented about how to design a cost calculation model for AM technology since it has not been industrialized to a significant extent. The purpose of this study is to develop an AM cost calculation model for the metal industry. The study is conducted through a qualitative case study at Seco Tools AB, a Swedish manufacturing company that carries out research and production operations of AM technology. By using literature review, data collection on the AM process, interviews with staff at the case company, a new AM cost calculation model is formulated specifically for the AM process at the case company. The proposed model is quantitatively and qualitatively validated with a set of real data from the case company and the satisfied result is obtained. The theoretical contribution of this study is the knowledge of what influencing factors should be included, what uncertainties exist and how corresponding adjustments should be made in a cost calculation model for AM. The practical contribution is the development of an innovative, user-friendly and generalizable cost model without user competence requirements within AM. Compared to the earlier models, the developed model is easier to apply when calculating the costs of a complex process. The model can be used as a basis for decision making in budgeting, scenario analysis, determination of occupancy rate or cost calculation of individual build jobs.Aktörer inom tillverkningsindustrin letar ständigt efter innovativa tillverkningsmetoder för att förbli konkurrenskraftiga. Additiv tillverkning (AM), även kallad 3D-printing, anses ha en god potential i utvecklingen mot industri 4.0. Även fast kostnadskontroll av AM-processer är ett viktigt steg mot framtida affärsmöjligheter, har lite forskning genomförts om hur man kan designa en kostnadsberäkningsmodell för AM-teknik eftersom den inte har industrialiserats i betydande utsträckning. Syftet med denna studie är att utveckla en kostnadsberäkningsmodell för AM inom metallindustrin. Studien genomförs genom en kvalitativ fallstudie på Seco Tools AB, ett svenskt tillverkningsföretag som bedriver forskning och produktionsverksamhet inom AM-teknik. Genom att använda litteraturinsamling, datainsamling om AM-processen samt intervjuer med personal på fallföretaget formas en ny kostnadsberäkningsmodell för AM specifikt mot AM-processen vid fallföretaget. Den föreslagna modellen är kvantitativt och kvalitativt validerad med en uppsättning verkliga data från fallföretaget och ett tillfredsställande resultat erhålls. Det teoretiska bidraget för denna studie är kunskapen om vilka påverkande faktorer som bör ingå, vilka osäkerheter som återfinns och hur motsvarande justeringar bör göras i en kostnadsberäkningsmodell för AM. Det praktiska bidraget är utvecklandet en innovativ, användarvänlig och generaliserbar kostnadsmodell utan kompetenskrav inom AM. Jämfört med tidigare modeller är den utvecklade modellen lättare att tillämpa vid beräkning av kostnader i en komplex process. Modellen kan användas som underlag för beslutsfattande vid budgetering, scenarioanalys, bestämning av beläggningsgrad eller kostnadsberäkning av enskilda byggjobb

Innovativ kostnadsberäkningsmodell för additiv tillverkning : En kvalitativ fallstudie med bidrag till utvecklingen av självkostnadsberäkning

Actors within the manufacturing industry are constantly looking for innovative manufacturing methods to remain competitive. Additive manufacturing (AM), also referred to as 3D printing, is considered to have a good potential in the development towards industry 4.0. Though the cost control of AM-processes is an important step towards future business opportunities, little research has been implemented about how to design a cost calculation model for AM technology since it has not been industrialized to a significant extent. The purpose of this study is to develop an AM cost calculation model for the metal industry. The study is conducted through a qualitative case study at Seco Tools AB, a Swedish manufacturing company that carries out research and production operations of AM technology. By using literature review, data collection on the AM process, interviews with staff at the case company, a new AM cost calculation model is formulated specifically for the AM process at the case company. The proposed model is quantitatively and qualitatively validated with a set of real data from the case company and the satisfied result is obtained. The theoretical contribution of this study is the knowledge of what influencing factors should be included, what uncertainties exist and how corresponding adjustments should be made in a cost calculation model for AM. The practical contribution is the development of an innovative, user-friendly and generalizable cost model without user competence requirements within AM. Compared to the earlier models, the developed model is easier to apply when calculating the costs of a complex process. The model can be used as a basis for decision making in budgeting, scenario analysis, determination of occupancy rate or cost calculation of individual build jobs.Aktörer inom tillverkningsindustrin letar ständigt efter innovativa tillverkningsmetoder för att förbli konkurrenskraftiga. Additiv tillverkning (AM), även kallad 3D-printing, anses ha en god potential i utvecklingen mot industri 4.0. Även fast kostnadskontroll av AM-processer är ett viktigt steg mot framtida affärsmöjligheter, har lite forskning genomförts om hur man kan designa en kostnadsberäkningsmodell för AM-teknik eftersom den inte har industrialiserats i betydande utsträckning. Syftet med denna studie är att utveckla en kostnadsberäkningsmodell för AM inom metallindustrin. Studien genomförs genom en kvalitativ fallstudie på Seco Tools AB, ett svenskt tillverkningsföretag som bedriver forskning och produktionsverksamhet inom AM-teknik. Genom att använda litteraturinsamling, datainsamling om AM-processen samt intervjuer med personal på fallföretaget formas en ny kostnadsberäkningsmodell för AM specifikt mot AM-processen vid fallföretaget. Den föreslagna modellen är kvantitativt och kvalitativt validerad med en uppsättning verkliga data från fallföretaget och ett tillfredsställande resultat erhålls. Det teoretiska bidraget för denna studie är kunskapen om vilka påverkande faktorer som bör ingå, vilka osäkerheter som återfinns och hur motsvarande justeringar bör göras i en kostnadsberäkningsmodell för AM. Det praktiska bidraget är utvecklandet en innovativ, användarvänlig och generaliserbar kostnadsmodell utan kompetenskrav inom AM. Jämfört med tidigare modeller är den utvecklade modellen lättare att tillämpa vid beräkning av kostnader i en komplex process. Modellen kan användas som underlag för beslutsfattande vid budgetering, scenarioanalys, bestämning av beläggningsgrad eller kostnadsberäkning av enskilda byggjobb