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Background: To inform future preventive measures including repeated vaccinations, we have searched for a clinically useful immune correlate of protection against fatal COVID-19 among nursing homes residents. Methods: We performed repeated capillary blood sampling with analysis of S-binding IgG in an open cohort of nursing home residents in Sweden. We analyzed immunological and registry data from 16 September 2021 to 31 August 2022 with follow-up of deaths to 30 September 2022. The study period included implementation of the 3rd and 4th mRNA monovalent vaccine doses and Omicron virus waves. Findings: A total of 3012 nursing home residents with median age 86 were enrolled. The 3rd mRNA dose elicited a 99-fold relative increase of S-binding IgG in blood and corresponding increase of neutralizing antibodies. The 4th mRNA vaccine dose boosted levels 3.8-fold. Half-life of S-binding IgG was 72 days. A total 528 residents acquired their first SARS-CoV-2 infection after the 3rd or the 4th vaccine dose and the associated 30-day mortality was 9.1%. We found no indication that levels of vaccine-induced antibodies protected against infection with Omicron VOCs. In contrast, the risk of death was inversely correlated to levels of S-directed IgG below the 20th percentile. The death risk plateaued at population average above the lower 35th percentile of S-binding IgG. Interpretation: In the absence of neutralizing antibodies that protect from infection, quantification of S-binding IgG post vaccination may be useful to identify the most vulnerable for fatal COVID-19 among the oldest and frailest. This information is of importance for future strategies to protect vulnerable populations against neutralization resistant variants of concern.

Microbial communities are the main drivers of biogeochemical cycling of multiple elements sustaining life in the ocean. The rapidity of their response to stressors and abrupt environmental changes implies that even fast and infrequent events can affect local transformations of organic matter and nutrients. Modern molecular techniques now allow for monitoring of microbial activities and functions in the environment through the analysis of genes and expressed genes contained in natural microbial assemblages. However, messenger RNA turnover in cells can be as short as 30 seconds and stability varies greatly between transcripts. Sampling of in situ communities involves an inevitable delay between the collection of seawater and the extraction of its RNA, leaving the bacterial communities plenty of time to alter their gene expression. The characteristics of microbial RNA turnover make time-series very difficult because samples need to be processed immediately to limit alterations to the metatranscriptomes. To address these challenges we designed an autonomous in situ fixation multi-sampler (AFISsys) for the reliable sampling of microbial metatranscriptomes at frequent intervals, for refined temporal resolution. To advance the development of this instrument, we examined the minimal seawater volume necessary for adequate coverage of community gene expression, and the suitability of phenol/ethanol fixation for immediate and long-term preservation of transcripts from a microbial community. We then evaluated the field eligibility of the instrument itself, with two case studies in a brackish system. AFISsys is able to collect, fix, and store water samples independently at a predefined temporal resolution. Phenol/ethanol fixation can conserve metatranscriptomes directly in the environment for up to a week, for later analysis in the laboratory. Thus, the AFISsys constitutes an invaluable tool for the integration of molecular functional analyses in environmental monitoring in brackish waters and in aquatic environments in general.

Early Neurological Deterioration (END) occur in up to 25% of patients with ischaemic stroke receiving stroke-unit-care and in 11-13.8% of patients treated with iv-tissue-Plasmniogen-Activator (iv-tPA). The aim of the study was to establish and compare the prevalence of END and symptomatic Intracranial Hemorrhage (sICH) in a prospectively designed registry of consecutive patients treated with iv-tPA to a historic cohort of iv-tPA eligible patients whom were hospitalized prior to implementation of iv-tPA-treatment but receiving otherwise comparable acute stroke care.Single center registry from a public Danish stroke-unit. Three-hundred-sixty-one unselected consecutive iv-tPA-treated patients admitted within 4.5 h from symptom-onset with symptoms of acute stroke and17 years of age. The iv-tPA-treated cohort was compared to a pre-tPA cohort of 246 iv-tPA-eligible patients who were admitted to the same stroke center from 1998 to 2001. Acute stroke care apart from iv-tPA was comparable. Outcome measures was assessed on admission and at 24 h; END as any increase in National Institute of Health Stroke Scale (NIHSS) and symptomatic Intracranial Hemorrhage (sICH) with use of CT-head-scan.END was observed in 27 (7.5%) of the 361 patients in the tPA-cohort and 43 (17.5%) of 246 in the pre-tPA-cohort, p 0.0001. Any ICH was detected in 23 (6.4%) and sICH in 3 (0.8%) of the iv-tPA-treated patients.END is significantly less frequent in acute stroke patients treated with iv-tPA. Deterioration due to ICH was rare and of limited severity in this population. END though remains a significant complication after stroke why more detailed knowledge on the various causes of END is needed to further improve patient outcome.

Controlling the growth of desired algal strains in microalgal-based wastewater treatment systems is challenging, and understanding the dynamics and interactions in the system is vital for proper management. A pilot-scale open photobioreactor located in Northern Sweden, was inoculated with the culture collection strain Scenedesmus dimorphus UTEX 417, which had been grown in Nordic climate for 8 years. The microbial diversity of eukaryotic and prokaryotic communities and their seasonal dynamics were revealed throughout the growth period by high-throughput sequencing of 18S and 16S rRNA genes and correlated with various environmental factors. S. dimorphus was stable in batch culture, but other microalgae appeared during semi-batch mode and co-dominated the system as a consequence of predation by zooplankton. Desmodesmus, Pseudocharaciopsis, Chlorella, Characium and Oocystis were the main microalgal species co-occurring in the system, with Characium and Chlorella acting as main competitors to Scenedesmus. Proteobacteria, Firmicutes, Bacteroidetes and Actinobacteria were the most abundant bacterial phyla, some of which showing significant positive or negative influence on Scenedesmus growth over time. Light, temperature, dissolved oxygen, pH and nutrients concentrations were found to have significant influence on the bacterial and/or zooplanktonic community changes.

Objective To assess the incidence of cancer in patients treated with growth hormone (GH) in KIGS—the Pfizer International Growth Database—without cancer or any other condition in medical history known to increase the risk of cancer. Study design Data were analyzed from patients with growth disorders enrolled in an observational survey KIGS who had no known increased risk of developing cancer before starting recombinant human GH treatment. The incidence of cancer in this patient cohort (overall, site-specific, and according to etiology of growth disorder) was compared with the incidence in the general population by using the standardized incidence ratio (ie, relating the observed to expected number of cases with stratification for age, sex, and country). Results A total of 32 new malignant neoplasms were reported in 58 603 patients, versus the 25.3 expected (incidence, 16.4 per 100 000 patient-years; standardized incidence ratio, 1.26; 95% confidence interval, 0.86-1.78). No category of growth disorder showed a statistically significant difference in observed compared with the expected number of cases. Conclusion There is no evidence in this series that GH treatment in young patients with growth disorders results in an increased risk of developing cancer relative to that expected in the normal population. However, surveillance for an extended time should continue to allow further assessment.

Articular cartilage is indispensable for joint function but has limited capacity for self-repair. Engineering of neocartilage in vitro is therefore a major target for autologous cartilage repair in arthritis. Previous analysis of neocartilage has targeted cellular organization and specific molecular components. However, the complexity of extracellular matrix (ECM) development in neocartilage has not been investigated by proteomics. To redress this, we developed a mouse neocartilage culture system that produces a cartilaginous ECM. Differential analysis of the tissue proteome of 3-week neocartilage and 3-day postnatal mouse cartilage using solubility-based protein fractionation targeted components involved in neocartilage development, including ECM maturation. Initially, SDS-PAGE analysis of sequential extracts revealed the transition in protein solubility from a high proportion of readily soluble (NaCl-extracted) proteins in juvenile cartilage to a high proportion of poorly soluble (guanidine hydrochloride-extracted) proteins in neocartilage. Label-free quantitative mass spectrometry (LTQ-Orbitrap) and statistical analysis were then used to filter three significant protein groups: proteins enriched according to extraction condition, proteins differentially abundant between juvenile cartilage and neocartilage, and proteins with differential solubility properties between the two tissue types. Classification of proteins differentially abundant between NaCl and guanidine hydrochloride extracts (n = 403) using bioinformatics revealed effective partitioning of readily soluble components from subunits of larger protein complexes. Proteins significantly enriched in neocartilage (n = 78) included proteins previously not reported or with unknown function in cartilage (integrin-binding protein DEL1; coiled-coil domain-containing protein 80; emilin-1 and pigment epithelium derived factor). Proteins with differential extractability between juvenile cartilage and neocartilage included ECM components (nidogen-2, perlecan, collagen VI, matrilin-3, tenascin and thrombospondin-1), and the relationship between protein extractability and ECM ultrastructural organization was supported by electron microscopy. Additionally, one guanidine extract-specific neocartilage protein, protease nexin-1, was confirmed by immunohistochemistry as a novel component of developing articular cartilage in vivo. The extraction profile and matrix-associated immunostaining implicates protease nexin-1 in cartilage development in vitro and in vivo.

The insulin-like growth factor (IGF) system exerts many effects on the growth and differentiation of both normal and malignant cells. The serum concentrations of insulin-like growth factor I (S-IGF-I), insulin-like growth factor-binding protein 2 (S-IGFBP-2) and insulin-like growth factor-binding protein 3 (S-IGFBP-3) and their inter-relations may differ in certain disease states from those seen in healthy individuals.To estimate age-, gender- and body mass index (BMI)-specific univariate, bivariate and trivariate 95% reference regions for these components in healthy adults and present indices that will facilitate interpretation of patient observations in relation to these reference regions.Blood samples were taken in the morning from 427 healthy, non-fasting German blood donors of both genders (age range, 18-79 years; BMI range, 16-44 kg/m(2)). Reference regions were developed with multivariate regression methods.Regression analyses showed that S-IGF-I and S-IGFBP-3 levels decrease with increasing age, whereas S-IGFBP-2 concentrations increase with age (P0.0001). Females had significantly higher S-IGFBP-3 levels than males (P0.0001) and similar S-IGF-I and S-IGFBP-2 concentrations. Increasing BMI was associated with decreasing S-IGFBP-2 (P0.0001), but was not significantly associated with the concentrations of the other two analytes. Controlling for age, gender and BMI, S-IGF-I and S-IGFBP-3 were positively correlated (r=0.58), whereas S-IGF-I and S-IGFBP-2, and S-IGFBP-2 and S-IGFBP-3 were negatively correlated (r=-0.11 and r=-0.18, respectively). Based on the regression models, which were controlled for age, gender and BMI, two- and three-dimensional 95% reference regions with associated patient indices were described for each pair of analytes and for the trio of analytes, respectively.The multivariate reference regions developed in this study should be useful in identifying patients with an abnormal IGF-IGFBP axis. It is suggested that introducing multidimensional reference regions and the described patient indices into clinical practice may aid monitoring of the safety of GH therapy. These patient indices may also be useful in the assessment of cancer risk.

Objective: The objective was to investigate the effects of 3 years of growth hormone (GH) replacement therapy in GH deficient (GHD) patients in Sweden. Design and patients: An open label study in 237 adults with GHD (116 men and 121 women), consecutively enrolled in KIMS (Pfizer’s international metabolic database) in Sweden. Measurements: QoL and healthcare consumption were determined using questionnaires [QoL-assessment of GHD in Adults (QoL-AGHDA), the psychological general well-being (PGWB) index and the patient life situation form (PLSF)]. Results: The mean starting dose of GH was 0.13 mg/day and the mean maintenance dose was 0.37 mg/day. The mean insulin-like growth factor I (IGF-I) SD score increased from −1.92 at baseline to 0.38 after 3 years. There was a sustained increase in QoL as measured by the QoL-AGHDA and PGWB questionnaires. The number of doctor visits and the number of days in hospital were reduced after 3 years of GH replacement. The number of days of sickleave decreased during the first 2 years of treatment, but returned towards baseline values after 3 years. Leisure-time physical activity and satisfaction with physical activity increased. Conclusion: Three years of GH replacement therapy induced a sustained improvement in QoL. Healthcare consumption was reduced, although the reduction in the number of days of sickleave was not statistically significant after 3 years of treatment.

Infrared (IR) intensities as determined by a combination of temperature dependent IR spectroscopy and temperature programmed desorption (TPD) were used to identify adsorption sites on Co and Co–Pd nano particles. It is shown that CO adsorption to metal atoms with a low metal coordination exhibits a strong intensity enhancement of the IR absorption as compared to regular surface sites. Density functional theory (DFT) calculations show that this intensity enhancement is due to a decoupling of the low coordinated metal atom bound to CO from those metal atoms embedded in the metal surface.

A model for bimetallic Co–Pd systems known to be active as Fischer–Tropsch catalysts has been constructed and studied under ultra-high vacuum conditions with respect to its adsorption and reaction properties. It was previously shown that subsequent deposition of Pd and Co results in the formation of core-shell structures; the morphology of the bimetallic particles can be controlled taking advantage of the different nucleation and growth behavior of Pd and Co. In this study, the reactive properties of pristine and oxidized bimetallic Co–Pd particles with O2, CO, D2 ,C 2D4, and C2H4 have been investigated using temperature programmed desorption and reaction spectroscopy. Hydrogen and CO adsorption is facile on the non-oxidized particles, but is severely inhibited on the oxidized particles. Ethylene binds to the non-oxidized particles in p-bonded and di-r-bonded configurations and dehydrogenates to form ethylidyne. In contrast, on the oxidized particles, only the p-bonded species is formed. To slightly different degrees, the bimetallic or monometallic particles are active for ethylene hydrogenation; the oxidized particles are essentially non-active for ethylene hydrogenation. � 2003 Elsevier B.V. All rights reserved.

Many commercially important mixtures contain complex polymers, e.g. paints and coatings. If a good thermodynamic description can be given of these systems it is possible to develop paints, which possess a certain set of properties and at the same time meet some basic requirement as, e.g. regarding the content of organic solvents. This work presents an investigation of the three polymer models Entropic-FV (EFV), UNIFAC-FV (UFV) and GC-Flory (GCF) for their capability of predicting solvent activity coefficients in binary systems containing complex polymers. It is possible to obtain good predictions at finite concentrations and satisfactory predictions at infinite dilution, particularly with the EFV model. The investigation shows that EFV is the most robust and stable of the models, which indicates that it is the most well suited model for further development of methods for predicting the miscibility behavior of paints and related systems.

Despite the growing evidence that plasma homocysteine is a cardiovascular risk factor, the mechanism behind the vascular injuries is still unknown. Studies are difficult as a result of the fact that little is known about the formation of different homocysteine species in vivo. Since extracellular glutathione and cysteine may influence the formation of different homocysteine species, we have in the present study investigated the different fractions of homocysteine and their relation to the different fractions of glutathione and cysteine in stroke patients and control subjects. We found a ratio of about 32–33% between reduced and total plasma glutathione concentrations and 2.6–3.0% between reduced and total plasma cysteine concentrations both in patients and in healthy control subjects. We noted an elevated concentration of total plasma homocysteine in stroke patients, but no difference in the ratio between reduced and total plasma homocysteine concentrations in patients and control subjects (mean value 1.20 and 1.10%, respectively). However, in a subgroup of patients with higher concentrations of total plasma homocysteine, we observed a significantly lower ratio of reduced to total plasma homocysteine compared to a subgroup of patients with lower concentration of total plasma homocysteine. A low reduced/total ratio of plasma homocysteine in combination with elevated plasma homocysteine concentrations might reflect an increased pro-oxidant activity in plasma from these patients. Thus, increased pro-oxidant activity in plasma might be one factor, besides genetic and nutritional factors, that could explain hyperhomocysteinemia. Since substantial evidence indicates that progression of atherosclerosis is related to enhanced pro-oxidant activity, the premature vascular disease associated with increased plasma homocysteine concentration might be as a result of increased pro-oxidant activity and the elevated plasma homocysteine concentration may only reflect the increased oxidative stress.

Despite the growing evidence that plasma homocysteine is a cardiovascular risk factor, the mechanism behind the vascular injuries is still unknown. In the present study we have investigated the possible role of hypomethylation as a cause of homocysteine-induced cell damage in two human cell lines. A significant growth retardation was observed in HeLa cell cultures in the combined presence of homocysteine and adenosine, but first at concentrations of 250 micromol/l of each. A significant decrease of intracellular glutathione concentration was noted both in the presence of homocysteine (250 micromol/l) alone and in the presence of the combination of homocysteine and adenosine (250 micromol/l). Intracellular concentration of homocysteine was increased to a similar extent both in the presence of homocysteine alone and in the presence of a combination of homocysteine and adenosine. Similar findings to those described for HeLa cell cultures were observed in endothelial cell cultures. Furthermore, in the presence of copper ions together with 100 micromol/l of adenosine and homocysteine a significantly retarded cell growth was observed in HeLa cell cultures. This finding shows that a combination of two potentially cell-damaging mechanisms (formation of oxygen radicals and hypomethylation) aggravated the retardation of cell growth compared to only one of these mechanisms being present. Thus, it is likely that several mechanisms of homocysteine-induced cell damage contribute to the increased rate of the atherogenic process observed in hyperhomocysteinemia.

Glutathione protects cells against oxidative damage, free radical damage and other types of toxicity. The aim of the present study was to investigate the impact on glutathione metabolism exerted by different thiol or redox reactive agents. Intracellular concentrations of glutathione in HeLa cell cultures were lowered after addition of agents mainly exerting oxidative stress (homocysteine and hydrogen peroxide), whereas thiol reactive oxidative agents (mercury ions, copper ions and hydroquinone) in concentrations not affecting cell growth seemed to stimulate the production of glutathione. Possibly, the thiol reactive agents decrease the concentration of glutathione, thereby stimulating further synthesis of glutathione, since glutathione synthesis is subject to feedback regulation by glutathione on γ-glutamylcysteine synthase. Redox changes after addition of thiol and redox reactive agents were also investigated. Copper ions lowered the concentrations of reduced forms of all extracellular thiols and of intracellular reduced cysteine in HeLa cell cultures. The addition of mercury ions, hydroquinone, homocysteine or hydrogen peroxide did not change the proportions between reduced and total thiol concentrations. After addition of buthionine sulfoxime, the total concentrations of intra- and extracellular glutathione were markedly decreased and the ratio between reduced and total glutathione concentrations was lowered. However, both cysteine and homocysteine exhibited normal ratios between the concentrations of reduced and total thiols in the presence of buthionine sulfoxime. This finding could be due to other cellular antioxidants, such as thioredoxin, ascorbic acid or tocopherols, maintaining redox status of these thiols.

Even mild hyperhomocysteinemia is associated with premature vascular disease. Despite the growing evidence that plasma homocysteine is a cardiovascular risk factor, the mechanism behind the vascular injuries is still unknown. Information about the metabolism of homocysteine is, therefore, essential for an understanding of its role in atherogenesis. In the present study we have, therefore, investigated the export mechanism of homocysteine. In HeLa cell lines the release of homocysteine was found to be a continuous process, which was increased in the presence of copper ions. High cell density led to a lowered release of homocysteine, probably due to a more extensive metabolism of the intracellular homocysteine. It was also found that HeLa cells were able to take up extracellularly released homocysteine and use it in the cellular metabolism. The ratio between intracellular homocysteine and the total amount of homocysteine is a measure of the ability of the cell to export the intracellularly produced homocysteine. The ratio also reflects the reuse of extracellular homocysteine. Under basal conditions, endothelial cells exported most of the intracellularly produced homocysteine and exhibited a very low concentration of homocysteine intracellularly, low reusage of exported homocysteine and consequently a low ratio in comparison with HeLa and hepatoma cell lines. After addition of homocysteine, all cell lines exhibited similar ratios. Thus, the intracellular homocysteine concentration in endothelial cells is more influenced by the extracellular concentration of homocysteine than is the intracellular concentration in HeLa and hepatoma cells. It may be speculated that this phenomenon could be associated with an increased sensitivity of endothelial cells to homocysteine and explain the association between hyperhomocysteinemia and vascular disease.

Different fractions (reduced and total) of thiols (homocysteine, cysteine and glutathione) were determined in HeLa cell cultures with and without addition of copper ions and/or homocysteine. In cell cultures without any addition the concentration of all intracellular thiols increased between 1 and 24 h of culture. Glutathione had the highest, whereas homocysteine showed the lowest, proportion of the reduced form. In the medium, there was a decrease of total cysteine during the incubation, but the amount of extracellular reduced cysteine increased. Both homocysteine and glutathione were released into the medium. The amount of exported homocysteine during the incubation exceeded several-fold the intracellular amount. There were no signs of cell toxicity induced by the high amounts of extracellularly added homocysteine (2000 mumol/l) in HeLa cell cultures, except a slight decrease in the concentration of intracellular glutathione. After the addition of copper ions (500 mumol/l) there was a retarded cell growth, decreased intracellular concentration of glutathione, increased release of glutathione into the medium and a lower proportion of all intra- and extracellular reduced thiols. After the addition of both copper ions and homocysteine to HeLa cell cultures, similar changes as with the addition of copper ions were noted except that the cell growth was still more retarded and that a very high level of intracellular homocysteine was noted at 1 h of incubation. N-acetylcysteine lowered, in these experiments, the intracellular accumulation of homocysteine and restored, to some extent, the cell growth. In an endothelial cell line even the presence of 500 mumol/l of homocysteine and 50 mumol/l of copper ions inhibited the cell growth and decreased the cellular level of glutathione. Whilst the levels of homocysteine in our short-time cell culture experiments are higher than the mild hyperhomocysteinemia thought to be atherogenic in humans (20-30 mumol/l), it is conceivable that over a longer time-course (several decades) these lower levels of homocysteine in the presence of copper ions could be sufficient to induce cellular effects similar to those found in the present study, eventually leading to atherosclerosis.

Ions of metals such as copper, mercury, silver and cadmium are known to exhibit a high affinity for thiol groups and may therefore severely disturb many metabolic functions in the cell. Copper ions are also known to catalyse the formation of toxic oxygen species through a series of redox reactions. In the present study, we have determined the concentration of reduced and total glutathione, cysteine and homocysteine in a cell culture system (HeLa cell line) after addition of these metal ions. The main findings of the metal ion effect on the total thiol concentrations are that all metal ions increased the release of glutathione into the medium. Since the intracellular concentration of glutathione did not decrease under these conditions, the synthesis of glutathione must have been increased. In contrast to the other metal ions, copper ions also increased the release of homocysteine into the medium, possibly through interaction with S-adenosylhomocysteine hydrolase. The main findings of metal ion effects on reduced thiol are that, at concentrations not interfering with cell growth, mercury, silver and cadmium ions increased the concentration of extracellular reduced glutathione, possibly reflecting the increase of total glutathione in the medium. In contrast to the other metal ions, the addition of even very low amounts of copper ions (1 mumol/l) decreased the concentration of intra- and extracellular reduced thiols indicating oxidative stress.

The Swedish experience of fatigue problems in large wind energy conversion systems (WECS) is summarized. The two prototypes, Nasudden and Maglarp, are discussed, with emphasis on blade-related topics for the steel and glass-fibre composite blades, respectively. The topics include loads, fatigue design, material properties, structural testing, inspection procedures and environmental effects. In-service fatigue-related experience during 5–6 years of operation is discussed in the light of these topics.