The present study describes diagnostic and prognostic abilities of Cerebrospinal fluid (CSF) Pentraxin 3 (PTX3) in central nervous system (CNS) infections. CSF PTX3 was measured retrospectively from ...174 patients admitted under suspicion of CNS infection. Medians, ROC curves and Youdens index was calculated. CSF PTX3 was significantly higher among all CNS infections and undetectable in most of the patients in the control group, and significantly higher in bacterial infections compared to viral and Lyme infections. No association was found between CSF PTX3 and Glasgow Outcome Score. PTX3 in the CSF can distinguish bacterial infection from viral and Lyme infections and non-CNS infections. Highest levels were found in bacterial meningitis. No prognostic abilities were found.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
To assess if SARS-CoV-2 (COVID-19) systemic disease can be determined by available nucleoprotein assays, we compared the performance of three commercial SARS-CoV-2 nucleoprotein (N) assays in plasma. ...A total of 272 plasma samples collected in the period November-December 2021 were analyzed by the methods Simoa SARS CoV-2 N Protein Advantage Kit Quanterix Simoa, Solsten SARS-CoV-2 Antigen enzyme immunosorbent assay (ELISA) Solsten ELISA, and Elecsys SARS-CoV-2 Antigen electrochemiluminescence immunoassay Elecsys ECLIA. Additionally, a dilution series of inactivated virus culture was analyzed by the three assays. The SARS CoV-2 PCR-status was not known for the patients. Linear correlation in the pairwise correlation between assays as well as linearity of dilution series of inactivated virus culture was estimated by Spearman score. Sensitivity and specificity were estimated by pairwise comparison. The three assays showed poor agreement on patient samples with regards to concentration. Performance on virus culture was excellent but with different level of detection (LOD). Positive vs negative results show comparable sensitivity and specificity of Quanterix Simoa and Solsten ELISA, with a higher LOD in Elecsys ECLIA and thus lower sensitivity and high specificity. N by all tested assays can be used as a marker for systemic COVID-19 disease.
Aims
To estimate the prevalence of gestational diabetes mellitus (GDM) in a Danish cohort comparing the current Danish versus the WHO2013 diagnostic criteria, and to evaluate adverse pregnancy ...outcomes among currently untreated women in the gap between the diagnostic thresholds.
Methods
Diagnostic testing was performed by a 75 g oral glucose tolerance test (OGTT) at 24–28 weeks’ gestation in a cohort of pregnant women. GDM diagnosis was based on the current Danish criterion (2-h glucose ≥ 9.0 mmol/L, GDM
DK
) and on the WHO2013 criteria (fasting ≥ 5.1, 1 h ≥ 10.0 or 2 h glucose ≥ 8.5 mmol/L, GDM
WHO2013
). Currently untreated women fulfilling the WHO2013 but not the Danish diagnostic criteria were defined as New-GDM-women (GDM
WHO2013
-positive and GDM
DK
-negative). Adverse outcomes risks were calculated using logistic regression.
Results
OGTT was completed by 465 women at a median of 25.7 weeks’ gestation. GDM
DK
prevalence was 2.2% (
N
= 10) and GDM
WHO2013
21.5% (
N
= 100). New-GDM was present in 19.4% (
N
= 90), of whom 90.0% had elevated fasting glucose. Pregnancies complicated by New-GDM had higher frequencies of pregnancy-induced hypertension (13.3% vs 4.1%,
p
= 0.002), large-for-gestational-age infants (22.2% vs 9.9%,
p
= 0.004), neonatal hypoglycaemia (8.9% vs 1.9%,
p
= 0.004) and neonatal intensive care unit admission (16.7% vs 5.8%,
p
= 0.002) compared to pregnancies without GDM.
Conclusions
GDM prevalence increased tenfold when applying WHO2013 criteria in a Danish population, mainly driven by higher fasting glucose levels. Untreated GDM in the gap between the current Danish and the WHO2013 diagnostic criteria resulted in higher risks of adverse pregnancy outcomes.
Serological biomarkers may be an option for early detection of colorectal cancer (CRC). The present study assessed eight cancer‐associated protein biomarkers in plasma from subjects undergoing first ...time ever colonoscopy due to symptoms attributable to colorectal neoplasia. Plasma AFP, CA19‐9, CEA, hs‐CRP, CyFra21‐1, Ferritin, Galectin‐3 and TIMP‐1 were determined in EDTA‐plasma using the Abbott ARCHITECT® automated immunoassay platform. Primary endpoints were detection of (i) CRC and high‐risk adenoma and (ii) CRC. Logistic regression was performed. Final reduced models were constructed selecting the four biomarkers with the highest likelihood scores. Subjects (N = 4,698) were consecutively included during 2010–2012. Colonoscopy detected 512 CRC patients, 319 colonic cancer and 193 rectal cancer. Extra colonic malignancies were detected in 177 patients, 689 had adenomas of which 399 were high‐risk, 1,342 had nonneoplastic bowell disease and 1,978 subjects had ‘clean’ colorectum. Univariable analysis demonstrated that all biomarkers were statistically significant. Multivariate logistic regression demonstrated that the blood‐based biomarkers in combination significantly predicted the endpoints. The reduced model resulted in the selection of CEA, hs‐CRP, CyFra21‐1 and Ferritin for the two endpoints; AUCs were 0.76 and 0.84, respectively. The postive predictive value at 90% sensitivity was 25% for endpoint 1 and the negative predictive value was 93%. For endpoint 2, the postive predictive value was 18% and the negative predictive value was 97%. Combinations of serological protein biomarkers provided a significant identification of subjects with high risk of the presence of colorectal neoplasia. The present set of biomarkers could become important adjunct in early detection of CRC.
What's new?
How can colorectal cancer be detected earlier? ‘Colonoscopies for everyone’ might be effective, but it is far too expensive. Instead, these authors propose a set of eight protein biomarkers that could signal the presence of cancer. They tested the blood‐based biomarker screening in patients who were already seeking a colonoscopy after experiencing bowel‐related symptoms. Of the nearly 4,700 subjects tested, colonoscopy uncovered 512 colorectal cancers and 399 high‐risk adenomas. The panel of biomarkers, in combination, successfully predicted the cancers. Next, the markers will need to be tested on asymptomatic subjects, to determine their usefulness as a general screening tool.
Background
Optimal balance between macro- and microcirculation in critically ill patients is crucial for ensuring optimal organ perfusion. Nitric oxide (NO) is a regulator of vascular hemostasis and ...tone. The availability of NO is controlled by asymmetric dimethylarginine (ADMA), symmetric dimethylarginine (SDMA), and the availability of the NO substrates arginine and homoarginine. We investigated the changes in plasma concentrations of ADMA, SDMA, arginine, and homoarginine days 1–5 of intensive care unit (ICU) admission and the association between the change in concentration days 1–3 and 30-day all-cause mortality.
Methods
Single-center cohort study of adult critically ill patients from the ICU at Copenhagen University Hospital – North Zealand. ADMA, SDMA, arginine, and homoarginine (NO-biomarkers) were measured on days 1–5. Initially, we determined the changes in NO-biomarkers days 1–5 with linear mixed models, and subsequently how the changes in NO-biomarkers days 1–3 were associated with 30-day all-cause mortality. Post-hoc we analyzed the association between plasma concentration at admission and 30-day all-cause mortality.
Results
In total 567 out of 577 patients had plasma samples from days 1–5. Plasma concentrations of ADMA and arginine increased from days 1–5. SDMA concentrations increased from days 1–2, followed by a decrease from days 2–5. Concentrations of homoarginine did not change from days 1–3 but slightly increased from days 3–5. In total 512 patients were alive 3 days after ICU admission. Among these patients, a daily twofold increase in ADMA concentration from days 1–3 was associated with decreased mortality in multivariate analysis (HR 0.45; 95% CI 0.21–0.98;
p
= 0.046). An increase in SDMA, arginine, or homoarginine was not associated with mortality. Post-hoc we found that a twofold increase in ADMA or SDMA concentrations at admission was associated with mortality (HR 1.78; 95% CI 1.24–2.57;
p
= 0.0025, and HR 1.41; 95% CI 1.05–1.90;
p
= 0.024, respectively).
Conclusions
Increasing ADMA concentrations on days 1–3 are inversely associated with mortality, however not with the same strength as high ADMA or SDMA concentrations at admission. We suggest that admission concentrations are the focus of future research on ADMA and SDMA as predictors of mortality or potential therapeutical targets in ICU patients.
The objective of the present feasibility study was to transfer single cell line cells to either microscopy slides for downstream immune characterization or to polymerase chain reaction tubes for ...downstream DNA quantitation. Tumour cell lines, SKBR3 and MCF7 and trophoblast cell line JEG-3 were spiked in healthy donor blood. The CytoTrack system was used to scan the spiked blood samples to identify target cells. Individual target cells were identified, picked by use of a CytoPicker and deposited to either a microscopic slide or a polymerase chain reaction tube (PCR). Single tumour cells on microscopic slides were further immunostained with human epidermal growth factor receptor 2 (Her2) and epithelial cell adhesion molecule (EpCAM). From the picked cells in polymerase chain reaction tubes, DNA was amplified, quantified and used for Short Tandem Repeat genotyping. Depositing rare cells to microscopy slides was laborious with only five cells per hour. In this study with a trained operator, the picked cells had an 80.5% recovery rate. Depositing single trophoblast cells in PCR tubes was a faster process with 10 cells in 5 min. Immunostaining of isolated cells by both Her2 and EpCAM was possible but showed varying staining intensity. Presence of trophoblasts and contaminating white blood cells in PCR tubes after cell picking was confirmed based on DNA yield and mixed Short Tandem Repeat profiles in five out of eight samples. Using the CytoPicker tool, single tumour and trophoblast cells were successfully isolated and moved from blood samples, allowing subsequent immunostaining or Short Tandem Repeat genotyping.
The collagens of the extracellular matrix are the most abundant structural proteins in the mammalian body. In tissue remodeling and in the invasive growth of malignant tumors, collagens constitute an ...important barrier, and consequently, the turnover of collagen is a rate-limiting process in these events. A recently discovered turnover route with importance for tumor growth involves intracellular collagen degradation and is governed by the collagen receptor, urokinase plasminogen activator receptor-associated protein (uPARAP or Endo180). The interplay between this mechanism and extracellular collagenolysis is not known. In this report, we demonstrate the existence of a new, composite collagen breakdown pathway. Thus, fibroblast-mediated collagen degradation proceeds preferentially as a sequential mechanism in which extracellular collagenolysis is followed by uPARAP/Endo180-mediated endocytosis of large collagen fragments. First, we show that collagen that has been pre-cleaved by a mammalian collagenase is taken up much more efficiently than intact, native collagen by uPARAP/Endo180-positive cells. Second, we demonstrate that this preference is governed by the acquisition of a gelatin-like structure by the collagen, occurring upon collagenase-mediated cleavage under native conditions. Third, we demonstrate that the growth of uPARAP/Endo180-deficient fibroblasts on a native collagen matrix leads to substantial extracellular accumulation of well defined collagen fragments, whereas, wild-type fibroblasts possess the ability to direct an organized and complete degradation sequence comprising both the initial cleavage, the endocytic uptake, and the intracellular breakdown of collagen.
1 The Copenhagen Muscle Research Centre, Department of Human Physiology, Institute of Exercise and Sport Sciences, University of Copenhagen, Copenhagen, Denmark; and 2 Diabetes Research Unit, ...Departments of Medicine, Physiology, and Biophysics, Section of Endocrinology, Boston University School of Medicine, Boston, Massachusetts
Submitted 16 August 2004
; accepted in final form 17 September 2004
Intracellular mechanisms regulating fat oxidation were investigated in human skeletal muscle during exercise. Eight young, healthy, moderately trained men performed bicycle exercise (60 min, 65% peak O 2 consumption) on two occasions, where they ingested either 1 ) a high-carbohydrate diet (H-CHO) or 2 ) a low-carbohydrate diet (L-CHO) before exercise to alter muscle glycogen content as well as to induce, respectively, low and high rates of fat oxidation. Leg fat oxidation was 122% higher during exercise in L-CHO than in H-CHO ( P < 0.001). In keeping with this, the activity of 2 -AMP-activated protein kinase ( 2 -AMPK) was increased twice as much in L-CHO as in H-CHO ( P < 0.01) at 60 min of exercise. However, acetyl-CoA carboxylase (ACC) Ser 221 phosphorylation was increased to the same extent (6-fold) under the two conditions. The concentration of malonyl-CoA was reduced 13% by exercise in both conditions ( P < 0.05). Pyruvate dehydrogenase activity was higher during exercise in H-CHO than in L-CHO ( P < 0.01). In H-CHO only, the concentrations of acetyl-CoA and acetylcarnitine were increased ( P < 0.001), and the concentration of free carnitine was decreased ( P < 0.01), by exercise. The data suggest that a decrease in the concentration of malonyl-CoA, secondary to 2 -AMPK activation and ACC inhibition (by phosphorylation), contributes to the increase in fat oxidation observed at the onset of exercise regardless of muscle glycogen levels. They also suggest that, with high muscle glycogen, the availability of free carnitine may limit fat oxidation during exercise, due to its increased use for acetylcarnitine formation.
acetylcarnitine; acetyl coenzyme A; pyruvate dehydrogenase activity; adenosine 5'-monosphosphate-activated protein kinase; acetyl coenzyme A carboxylase
Address for reprint requests and other correspondence: C. Roepstorff, The Copenhagen Muscle Research Centre, Institute of Exercise and Sport Sciences, Dept. of Human Physiology, Universitetsparken 13, DK-2100 Copenhagen Ø, Denmark (E-mail: croepstorff{at}ifi.ku.dk )
Previous studies have described the magnitude and time course by which several genes are regu‐lated within exercising skeletal muscle. These include interleukin‐6 (IL‐6), interleukin‐8 (IL‐8), heme ...oxygen‐ase‐1 (HO‐1), and heat shock protein‐72 (HSP72), which are involved in secondary signaling and preservation of intracellular environment. However, the primary signaling mechanisms coupling contraction to transcription are unknown. We hypothesized that exercise‐induced nitric oxide (NO) production is an important signaling event for IL‐6, IL‐8, HO‐1, and HSP72 expression in muscle. Twenty healthy males participated in the study. By realtime PCR, mRNA levels for 11 genes were determined in thigh muscle biopsies obtained 1) before and after 2 h knee extensor exercise without (control) and with con‐comitant NO synthase inhibition (nitro‐L‐arginine methyl ester, L‐NAME, 5 mgkg_1);or 2) before and after 2 h femoral artery infusion of the NO donor nitroglycerin (NTG, 1.5 μgkg_1min_1). L‐NAME caused marked reductions in exercise‐induced expression of 4 of 11 mRNAs including IL‐6, IL‐8, and HO‐1. IL‐6 protein release from the study leg to the circulation increased in the control but not in the L‐NAME trial. NTG infusion significantly augmented expression of the mRNAs attenuated by L‐NAME. These findings advance the novel concept that NO production contributes to regulation of gene expression in muscle during exercise. Subsequently, we sought evidence for involvement of AMP‐activated kinase or nuclear factor kappa B, but found none.—Steensberg, A., Keller, C., Hillig, T., Frosig, C., Wojtaszewski, J. F. P., Pedersen, B. K., Pilegaard, H., Sander, M. Nitric oxide production is a proximal signaling event controlling exercise‐induced mRNA expression in human skeletal muscle. FASEB J. 21, 2683–2694 (2007)
The urokinase receptor, urokinase receptor (uPAR), is a glycosylphosphatidylinositol-anchored membrane protein engaged in pericellular proteolysis and cellular adhesion, migration, and modulation of ...cell morphology. A direct matrix adhesion is mediated through the binding of uPAR to vitronectin, and this event is followed by downstream effects including changes in the cytoskeletal organization. However, it remains unclear whether the adhesion through uPAR-vitronectin is the only event capable of initiating these morphological rearrangements or whether lateral interactions between uPAR and integrins can induce the same response. In this report, we show that both of these triggering mechanisms can be operative and that uPAR-dependent modulation of cell morphology can indeed occur independently of a direct vitronectin binding. Expression of wild-type uPAR on HEK293 cells led to pronounced vitronectin adhesion and cytoskeletal rearrangements, whereas a mutant uPAR, uPARW32A with defective vitronectin binding, failed to induce both phenomena. However, upon saturation of uPARW32A with the protease ligand, pro-uPA, or its receptor-binding domain, the ability to induce cytoskeletal rearrangements was restored, although this did not rescue the uPAR-vitronectin binding and adhesion capability. On the other hand, using other uPAR variants, we could show that uPAR-vitronectin adhesion is indeed capable and sufficient to induce the same morphological rearrangements. This was shown with cells expressing a different single-site mutant, uPARY57A, in the presence of a synthetic uPAR-binding peptide, as well as with wild-type uPAR, which underwent cytoskeletal rearrangements even when cultivated in uPA-deficient serum. Blocking of integrins with an Arg-Gly-Asp-containing peptide counteracted the matrix contacts necessary to initiate the uPAR-dependent cytoskeletal rearrangements, whereas inactivation of the Rac signaling pathway in all cases suppressed the occurrence of the same events.
PDF
