Alpha-1-acid glycoprotein (AGP) is an acute phase glycoprotein in blood, which is primarily synthetized in the liver and whose biological role is not completely understood. It consists of 45% ...carbohydrates that are present in the form of five N-linked complex glycans. AGP N-glycosylation was shown to be changed in many different diseases, and some changes appear to be disease-specific; thus, it has a great diagnostic and prognostic potential. However, AGP glycosylation was mainly analyzed in small cohorts and without detailed site-specific glycan information. Here, we developed a cost-effective method for a high-throughput and site-specific N-glycosylation LC-MS analysis of AGP which can be applied on large cohorts, aid in search for novel disease biomarkers, and enable better understanding of AGP’s role and function in health and disease. The method does not require isolation of AGP with antibodies and affinity chromatography, but AGP is enriched by acid precipitation from 5 μl of bloodplasma in a 96-well format. After trypsinization, AGP glycopeptides are purified using a hydrophilic interaction chromatography-based solid-phase extraction and analyzed by reversed-phase-liquid chromatography-electrospray ionization-MS. We used our method to show for the first time that AGP N-glycan profile is stable in healthy individuals (14 individuals in three time points), which is a requirement for evaluation of its diagnostic potential. Furthermore, we tested our method on a population including individuals with registered hyperglycemia in critical illness (59 cases and 49 controls), which represents a significantly increased risk of developing type 2 diabetes. Individuals at higher risk of diabetes presented increased N-glycan branching on AGP’s second glycosylation site and lower sialylation of N-glycans on AGP’s third and AGP1’s fourth glycosylation site. Although this should be confirmed on a larger prospective cohort, it indicates that site-specific AGP N-glycan profile could help distinguish individuals who are at risk of type 2 diabetes.
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•Cost-effective method for high-throughput detailed AGP glycoprofiling is presented.•Site-specific AGP N-glycan profile can be obtained from 5 μl of blood plasma.•AGP N-glycan profile is stable in a healthy individual.•AGP glycoprofile could help identify individuals who are at risk of type 2 diabetes.
For the first time, a cost-effective method for a high-throughput detailed AGP N-glycosylation profiling was developed, which includes site-specific glycosylation information. Using the method, it was demonstrated that AGP N-glycan profile is stable in a healthy individual. Furthermore, using the method on a pilot cohort, it was found that site-specific AGP N-glycan profile could help distinguish individuals who are at risk of type 2 diabetes. The method presents a new valuable tool for investigation of AGP’s large biomarker potential.
BackgroundPulmonary embolism (PE) is a common diagnosis in an emergency department. It also represents a large share of patients admitted to hospital wards. Patients with PE can be risk-stratified ...and discharged early from the emergency department. This results in better availability of hospital beds for other patients and a significant reduction of treatment costs for the healthcare system. This paper aims to describe the protocols used in our emergency department, with special emphasis on risk stratification, for adverse events and bleeding risk, treatment strategies, and outcomes for this type of protocol. Materials and methodsThis paper is a retrospective analysis of patients discharged from the emergency department in a of two-year period (2020-2021) with a low-risk pulmonary embolism. ResultsWe have included in this study 42 patients discharged after a short-term observation from the emergency department (<24h) or short-term hospitalization; <24h). Ninety-one percent of patients were discharged with direct oral anticoagulant as a treatment for PE. We did not notice any adverse events (hemorrhage, progression of PE, or major cardiovascular issues). ConclusionIn the cohort of patients with PE, early discharge and outpatient treatment was safe and effective, with lower healthcare costs and almost no adverse events for patients.
Cell-free DNA has been associated with outcome in several acute conditions including two reports concerning the outcomes after cardiac arrest that found association of circulating DNA quantities at ...admission with mortality. The origins of cell-free DNA are primarily necrosis and apoptosis, which in cardiac arrest occur during ischaemia (“no-flow” and “low-flow” period), during reperfusion injury and as a consequence of post-arrest inflammatory response. Respecting the facts that significant cellular damage may occur during the post-arrest period, and that damage might be reduced by mild therapeutic hypothermia, we investigated the prognostic value of cell free DNA at ICU admission and 24h after admission.
A prospective study was conducted in three university associated intensive care units and included patients resuscitated from non-traumatic out-of-hospital cardiac arrest. Patient data were collected in accordance with the Utstein protocol. Therapeutic hypothermia was performed according to ICU policies.
Blood for cell-free DNA quantification was sampled at admission and at 24±1h after admission. Outcome measures were hospital morality and cerebral performance expressed with CPC scale at discharge.
Inclusion criteria were met in 67 patients; 24-h mortality was 37.3% and hospital mortality was 71.6%. The following variables were associated with 24-h mortality in univariate analysis: asystole as the presenting rhythm, “no-flow” time, “low-flow” time and cell-free DNA at admission (median 0.081 in survivors vs. 0.160ng/μl in non-survivors; P=0.038). Multivariate analysis that included the above variables showed that no-flow time and low-flow time were independently associated with 24-h mortality.
Hospital mortality was associated with the following factors: “low flow” time, coronary intervention, cell-free DNA at ICU admission and at 24h after admission (0.042 vs. 0.188ng/μl; P=0.048). ROC curve for cell-free DNA 24h post-admission showed sensitivity of 81.0% and specificity of 78.3% for the cut-off value of 0.115ng/μl.
Multivariate analysis showed that “low-flow” time and cell-free DNA at 24h after ICU admission were independently associated with hospital mortality. Cell free DNA showed different dynamics in patients who were and who were not treated with mild therapeutic hypothermia: it decreased in treated patients and slightly increased in non-treated patients.
Cell-free DNA quantity at ICU admission and 24h after admission is associated with hospital mortality. Further studies will need to additionally investigate possible practical use of this new laboratory marker in patients resuscitated from cardiac arrest.
Objective and Design To investigate the association between the severity of sepsis and changes in sialylation of serum proteins we have conducted a single center pilot study. Subjects and Methods ...Sialylation of transferrin (with enzyme-linked lectin assay-ELLA) and total serum proteins (with colorimetric assay) as well as serum iron and transferrin levels were measured in 27 patients with sepsis through the first eight days of the disease. Results Total serum sialylation increased in the first two days, transferrin sialylation decreased, while serum iron and transferrin fell. Patients who developed severe sepsis had either a small or marked change in transferrin sialylation while in patients with mild sepsis sialylation decreased moderately. Conclusion We hypothesize that the change in transferrin sialylation could be a reflection of the intensity of inflammatory response which is insufficient if under-expressed and detrimental if over-expressed. This new feature is a potential marker of sepsis severity early in the disease.
Background
A previous National Institute for Health and Care Research study Harrison DA, Ferrando-Vivas P, Shahin J, Rowan KM. Ensuring comparisons of health-care providers are fair: development and ...validation of risk prediction models for critically ill patients.
Health Serv Deliv Res
2015;
3
(41) identified the need for more research to understand risk factors and consequences of critical care and subsequent outcomes.
Objectives
First, to improve risk models for adult general critical care by developing models for mortality at fixed time points and time-to-event outcomes, end-stage renal disease, type 2 diabetes, health-care utilisation and costs. Second, to improve risk models for cardiothoracic critical care by enhancing risk factor data and developing models for longer-term mortality. Third, to improve risk models for in-hospital cardiac arrest by enhancing risk factor data and developing models for longer-term mortality and critical care utilisation.
Design
Risk modelling study linking existing data.
Setting
NHS adult critical care units and acute hospitals in England.
Participants
Patients admitted to an adult critical care unit or experiencing an in-hospital cardiac arrest.
Interventions
None.
Main outcome measures
Mortality at hospital discharge, 30 days, 90 days and 1 year following critical care unit admission; mortality at 1 year following discharge from acute hospital; new diagnosis of end-stage renal disease or type 2 diabetes; hospital resource use and costs; return of spontaneous circulation sustained for > 20 minutes; survival to hospital discharge and 1 year; and length of stay in critical care following in-hospital cardiac arrest.
Data sources
Case Mix Programme, National Cardiac Arrest Audit, UK Renal Registry, National Diabetes Audit, National Adult Cardiac Surgery Audit, Hospital Episode Statistics and Office for National Statistics.
Results
Data were linked for 965,576 critical care admissions between 1 April 2009 and 31 March 2016, and 83,939 in-hospital cardiac arrests between 1 April 2011 and 31 March 2016. For admissions to adult critical care units, models for 30-day mortality had similar predictors and performance to those for hospital mortality and did not reduce heterogeneity. Models for longer-term outcomes reflected increasing importance of chronic over acute predictors. New models for end-stage renal disease and diabetes will allow benchmarking of critical care units against these important outcomes and identification of patients requiring enhanced follow-up. The strongest predictors of health-care costs were prior hospitalisation, prior dependency and chronic conditions. Adding pre- and intra-operative risk factors to models for cardiothoracic critical care gave little improvement in performance. Adding comorbidities to models for in-hospital cardiac arrest provided modest improvements but were of greater importance for longer-term outcomes.
Limitations
Delays in obtaining linked data resulted in the data used being 5 years old at the point of publication: models will already require recalibration.
Conclusions
Data linkage provided enhancements to the risk models underpinning national clinical audits in the form of additional predictors and novel outcomes measures. The new models developed in this report may assist in providing objective estimates of potential outcomes to patients and their families.
Future work
(1) Develop and test care pathways for recovery following critical illness targeted at those with the greatest need; (2) explore other relevant data sources for longer-term outcomes; (3) widen data linkage for resource use and costs to primary care, outpatient and emergency department data.
Study registration
This study is registered as NCT02454257.
Funding details
This project was funded by the National Institute for Health and Care Research (NIHR) Health and Social Care Delivery Research programme and will be published in full in
Health and Social Care Delivery Research
; Vol. 10, No. 39. See the NIHR Journals Library website for further project information.
Early prediction of severe acute pancreatitis is one of the problems in clinical practice. Since many diseases are associated with alteration in glycosylation, in this work we studied sialylation of ...transferrin and serum proteins in acute pancreatitis.
Sialylation was analyzed during first eight days of hospitalization of 30 patients and compared to 28 healthy controls. Transferrin sialylation was measured using enzyme linked lectin assay, while sialic acid on proteins was measured using resorcinol method.
Both analyzed parameters changed during studied period. The change in transferrin sialylation between Day1 and Day2 of hospitalization was shown to be an early prognostic marker of acute pancreatitis, with better sensitivity (88.9%) and specificity (90.5%) than other markers tested.
Sialylation of transferrin and total serum proteins reflects the intensity of inflammatory response during acute pancreatitis and could be used as prognostic parameter for disease severity.
Background: Pulmonary embolism (PE) is a common diagnosis in an emergency department. It also represents a large share of patients admitted to hospital wards. Patients with PE can be risk-stratified ...and discharged early from the emergency department. This results in better availability of hospital beds for other patients and a significant reduction of treatment costs for the healthcare system. This paper aims to describe the protocols used in our emergency department, with special emphasis on risk stratification, for adverse events and bleeding risk, treatment strategies, and outcomes for this type of protocol. Materials and methods: This paper is a retrospective analysis of patients discharged from the emergency department in a of two-year period (2020-2021) with a low-risk pulmonary embolism. Results: We have included in this study 42 patients discharged after a short-term observation from the emergency department (<24h) or short-term hospitalization; <24h). Ninety-one percent of patients were discharged with direct oral anticoagulant as a treatment for PE. We did not notice any adverse events (hemorrhage, progression of PE, or major cardiovascular issues). Conclusion: In the cohort of patients with PE, early discharge and outpatient treatment was safe and effective, with lower healthcare costs and almost no adverse events for patients Key words: pulmonary embolism, emergency department, early discharge, outpatient treatment Uvod: Plucna embolija (PE) predstavlja jednu od cestih dijagnoza u svakodnevnom radu hitnog prijema i dovodi do velikog broja prijema u bolnicu, sto nije uvijek nuzno. Posto se pacijenti na temelju obrade i stratifikacije rizika mogu obraditi i otpustiti iz hitnog prijema, to moze dovesti do rasterecenja bolnickih odjela, te smanjenja troskova u sustavu zdravstva. Cilj ovog rada je opisati protokol koji se koristi u nasem centru, koji ukljucuje obradu stratifikaciju rizika za kompliciranu klinicku sliku i krvarenje, terapijske opcije i nezeljene dogadaje ovakve vrste lijecenja Materijali i metode: Ovo je retrospektivna analiza pacijenata koji su u razdoblju od dvije godine (2020 i 2021 godina) otpusteni iz hitnog prijema kao plucna embolija niskog rizika. Rezultati: U studiju je ukljuceno 42 pacijenata koja su otpustena iz OHBP KBC Zagreb nakon kratkotrajne opservacije (do 24h) ili su otpustena iz bolnice nakon kratkotrajne hospitalizacije (do 24h). U 91% slucaja je pacijentu u terapiju uveden direktni oralni antikoagulans. Kod niti jednog pacijenta nije bilo nezeljenih dogadaja (krvarenje, progresija tromboze ili veliki kardiovaskularni incident) Zakljucak: U ovoj kohorti jasno se vidi da je rani otpust pacijenata sa plucnom embolijom niskog rizika siguran nacin za lijecenje ove bolesti, te da dovodi do veceg zadovoljstva pacijenata, manjeg boja hospitalizacija, manjih troskova za bolnice, te zanemarivog broja nezeljenih ucinaka lijecenja. Kljucne rijeci: plucna embolija, hitni prijem, izvanbolnicko lijecenje, rani otpust
Statini, inhibitori hidroksimetilglutaril koenzim A reduktaze, temelj su liječenja hiperkolesterolemije u kardiovaskularnih bolesnika. Osim učinka na sintezu kolesterola, imaju brojne druge učinke. ...Pleiotropni učinci statina su protu-upalni, imunomodulatorni, antioksidantni, antitrombotski i endotel-stabilizirajući. Svi spomenuti učinci mogli bi imati povoljne posljedice tijekom sistemskog odgovora na infekciju, tj. u sepsi, ali i u drugim sistemskim upalnim stanjima. Dan je pregled farmakoloških učinaka statina i pregled studija koje su ispitivale učinke statina u akutnom upalnom odgovoru i sepsi.
Chromic acid is a strong metal acid and acute poisoning is very rare, but very serious with severe skin injury, renal and liver failure. The majority of published cases were suicide attempts with ...lethal outcomes. We describe the case of a 55-year-old man who had accidentally taken a sip of 20% chromic acid (estimated chromium intake: 2.3 g). Renal and liver failure were not present at presentation, but appeared later in the course of disease. He was treated with hemodialysis, no chelating agents or other methods for enhancing elimination were used. Liver and renal function improved over the next 30 days and the patient was discharged after 45 days of hospitalization with no need for dialysis. In the follow-up period of eight months his renal function remained depressed, but stable.
Aims/hypothesis
Better understanding of type 2 diabetes and its prevention is a pressing need. Changes in human plasma N-glycome are associated with many diseases and represent promising diagnostic ...and prognostic biomarkers. Variations in glucose metabolism directly affect glycosylation through the hexosamine pathway but studies of plasma glycome in type 2 diabetes are scarce. The aim of this study was to determine whether plasma protein N-glycome is changed in individuals who are at greater risk of developing type 2 diabetes.
Methods
Using a chromatographic approach, we analysed N-linked glycans from plasma proteins in two populations comprising individuals with registered hyperglycaemia during critical illness (increased risk for development of type 2 diabetes) and individuals who stayed normoglycaemic during the same condition: AcuteInflammation (59 cases vs 49 controls) and AcuteInflammation Replication (52 cases vs 14 controls) populations. N-glycome was also studied in individuals from FinRisk (37 incident cases of type 2 diabetes collected at baseline vs 37 controls), Orkney Complex Disease Study (ORCADES; 94 individuals with HbA
1c
> 6.5% 47.5 mmol/mol vs 658 controls) and Southall and Brent Revisited (SABRE) cohort studies (307 individuals with HbA
1c
> 6.5% 47.5 mmol/mol vs 307 controls).
Results
Individuals with increased risk for diabetes type 2 development (AcuteInflammation and AcuteInflammation Replication populations), incident cases of type 2 diabetes collected at baseline (FinRisk population) and individuals with elevated HbA
1c
(ORCADES and SABRE populations) all presented with increased branching, galactosylation and sialylation of plasma protein N-glycans and these changes were of similar magnitude.
Conclusions/interpretation
Increased complexity of plasma N-glycan structures is associated with higher risk of developing type 2 diabetes and poorer regulation of blood glucose levels. Although further research is needed, this finding could offer a potential new approach for improvement in prevention of diabetes and its complications.
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