High-Throughput and Site-Specific N-Glycosylation Analysis of Human Alpha-1-Acid Glycoprotein Offers a Great Potential for New Biomarker Discovery.

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.

Successful treatment of late obstetric ethylene glycol intoxication with ethanol via the enteral route: a case report.

Late obstetric ethylene glycol intoxication represents a diagnostic challenge for acute care physicians and an impending life threat with life-long implications for both the mother and the fetus. The metabolism of ethylene glycol to its toxic metabolites during late pregnancy is unpredictable due to maternal physiological changes. Namely, the hallmark signs and symptoms of ethylene glycol intoxication can mimic those of late pregnancy-related high blood pressure disorders, which makes it difficult to correctly diagnose the condition. Therefore, it is crucial to promptly recognize late obstetric ethylene glycol intoxication and initiate specific treatment, but evidence-based recommendations are not available to guide its most effective emergent treatment. We present our department's emergent management of late-obstetric ethylene glycol intoxication. The parturient was stabilized by inhibiting ethylene glycol metabolism, alongside general supportive care measures. The enhancement of its toxic metabolites was eliminated by administering ethanol via the enteral route, which progressively improved the parturient's clinical course and led to the on-term delivery of a healthy child. Our case shows the importance of a meticulous emergent assessment, prompt diagnosis, and carefully planned multidisciplinary treatment in the emergency department in improving outcomes after ethylene glycol intoxication in late pregnancy.

Outpatient treatment of pneumonia in a setting with and without an infectious disease doctor.

AIM: To compare the outpatient treatment of community acquired pneumonia (CAP) by infectious disease doctors (IDDs) and doctors of other specialties (nIDDs). METHODS: We retrospectively identified 600 outpatients with CAP: 300 treated by IDDs and 300 by nIDDs in two tertiary hospitals during 2019. The two groups were compared in terms of adherence to guidelines, antibiotic group prescription, frequency of combined treatment, and treatment duration. RESULTS: IDDs prescribed significantly more first-line treatment (P<0.001) and alternative treatment (P=0.008). NIDDs prescribed more reasonable (P<0.001) and unnecessary (P=0.002) second-line treatment, and inadequate treatment (P=0.004). IDDs significantly more frequently prescribed amoxicillin (P<0.001) for typical and doxycycline (P=0.045) for atypical CAP, while nIDDs significantly more frequently prescribed amoxicillin-clavulanate (P<0.001) for typical and fluoroquinolones for both typical (P<0.001) and atypical (P<0.001) CAP. No significant differences were found in the frequency of combined treatment, which exceeded 50% in both groups, or in treatment duration. CONCLUSIONS: Outpatient treatment of CAP in the absence of IDDs meant more broad-spectrum antibiotic prescription and more disregard for national guidelines. Our results highlight the need for antibiotic stewardship, especially in settings with no IDDs.

OUTPATIENT TREATMENT OF PULMONARY EMBOLISM - A SINGLE-CENTER EXPERIENCE.

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.

Correction to: Increased plasma N-glycome complexity is associated with higher risk of type 2 diabetes.

The authors regret that Nish Chaturvedi's name was spelt incorrectly in the author list. The details given in this correction are correct.

Increased plasma N-glycome complexity is associated with higher risk of type 2 diabetes.

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 HbA1c > 6.5% [47.5 mmol/mol] vs 658 controls) and Southall and Brent Revisited (SABRE) cohort studies (307 individuals with HbA1c > 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 HbA1c (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.

Severe acute pancreatitis as a part of multiple dysfunction syndrome.

Acute pancreatitis is a disease with various degrees of clinical manifestations. Mild and moderate severe acute pancreatitis is an illness characterized with chemical inflammation which, in general, passes without major complications. Clinical picture of severe acute pancreatitis other side is commonly complicated with functional deterioration of other organs, and frequently has characteristics of multiple organ dysfunction or failure syndrome with or without bacterial super infection. We studied 82 patients admitted to the intensive care unit with severe acute pancreatitis, 14 died. The mortality was in statistically significant correlation with the severity of clinical condition at admission assessed by APACHE II score, and higher Ranson's and Glasgow criteria by admission. Adequate volume supplementation, on time, as well as percutaneous drainage of infected pancreas collection reduces a risk of pure outcome.

[Guidelines for the management of hyperglycaemia in hospitalised adult patients]. / Smjernice za zbrinjavanje hiperglikemije u odraslih hospitaliziranih bolesnika.

INTRODUCTION: The aim was to prepare guidelines for the management of in-hospital hyperglycaemia in adult patients in intensive care units and regular wards. CONTRIBUTORS: Working group led by two coordinators consists of repre- sentatives of professional societies within the Croatian Medical Association. EVIDENCE: These guidelines are derived from the guidelines of international professional societies. Level of evidence and strength of recommendation are evaluated according to GRADE (Grading of Recommendations, Assessment, Development and Evaluation) system. Decision making process: Working group produced Guidelines draft by a consensus. The draft has been delivered to professional societies for review. Final document is accepted by all included societies. CONCLUSION: Hyperglycaemia is a frequent, serious and demanding complication in hospitalized patients. Results of published studies suggest that its regulation decreases morbidity and mortality. Implementation of locally developed standardized protocols promoting basal-bolus insulin regimen is regarded as the most important measure for management of hyperglycaemia in hospital. Present guidelines are a set of practical, rational and feasible recommendations and suggestions.

Exploratory analysis of glial fibrillary acidic protein and ubiquitin C-terminal hydrolase L1 in management of patients with mild neurological symptoms undergoing head computed tomography scan at the emergency department: a pilot study from a Croatian tertiary hospital.

BACKGROUND: Diagnostic accuracy of glial fibrillary acidic protein (GFAP) and ubiquitin C-terminal hydrolase L1 (UCH-L1) in identification of intracranial abnormalities detected by computed tomography (CT) in mild traumatic brain injury (mTBI), and in patients with mild neurological symptoms not caused by head trauma but suspected with a neurological disorder, was examined. METHODS: GFAP and UCH-L1 were determined using the chemiluminescence immunoassays on the Alinity i analyzer (Abbott Laboratories). RESULTS: Significantly higher GFAP (median 53.8 vs 25.7 ng/L, P < .001) and UCH-L1 (median 350.9 vs 153.9 ng/L, P < .001) were found in mTBI compared to non-head trauma patients. In mTBI diagnostic sensitivity (Se) and specificity (Sp) for the combination of GFAP and UCH-L1 were 100% and 30.9%, respectively, with area under the curve (AUC) 0.655. GFAP alone yielded Se 85.7%, Sp 41.8%, and AUC 0.638, while UCH-L1 yielded Se 57.1%, Sp 56.4%, and AUC 0.568. In non-head trauma patients, the combination of GFAP and UCH-L1 showed Se 100%, Sp 87.9%, and AUC 0.939, while GFAP alone demonstrated Se 100%, Sp 90.9%, and AUC 0.955. CONCLUSIONS: If these results are reproduced on a larger sample, GFAP and UCH-L1 may reduce CT use in patients with mild neurological symptoms after systemic causes exclusion and neurologist's evaluation.

Prior statin therapy is associated with milder course and better outcome in acute pancreatitis--a cohort study.

BACKGROUND: Statin treatment was shown to be associated with improved outcomes in several inflammatory conditions. We wanted to evaluate the effects of statin therapy on the course and outcome of acute pancreatitis (AP). METHODS: A prospective cohort study included patients with acute pancreatitis divided into two groups according to statin use prior to hospitalization. Age, sex, etiology of AP, Ranson's score, APACHE II score and maximal CRP were recorded. Outcome measures were hospital length of stay and mortality. Matching of patients for matched analyses was done using individual matching and propensity score matching using variables a priori associated with course and outcome of acute pancreatitis. RESULTS: Inclusion criteria were met for 1062 patients of whom 92 were taking statins. Statin users were older and had higher body mass indexes. Severe disease was more common in the no-statin group than in statin group (20.6% vs. 8.7% respectively). All severity markers were also higher in the no-statin group. All cause mortality was not different, while cardiovascular mortality was higher in the statin group in the cohort analysis. After matching by either method, the severity of disease was greater for the patients without statins treatment. Pancreatitis related mortality was higher in the no-statin group after matching. Among patients who developed severe AP, statin users showed lower Ranson's and APACHE II scores and lower maximal CRP. CONCLUSIONS: Prior statin treatment significantly reduces morbidity and mortality in acute pancreatitis. Further studies are needed to evaluate possible therapeutic use of statins in acute pancreatitis.

Haemoglobin A1c-based screening for prediabetes and diabetes mellitus: a multi-center study in Croatian adult population.

INTRODUCTION: Based on the hypothesis that there is a substantial rate of adults with prediabetes and undiagnosed diabetes mellitus (DM), our aim was to perform haemoglobin A1c (HbA1c)-based screening in a cohort of Croatian adults and estimate the prevalence of prediabetes and undiagnosed DM according to American Diabetes Association criteria. MATERIALS AND METHODS: This multi-center, cross-sectional study performed in six Croatian hospitals included 5527 patients aged 40 to 70 years admitted to the Emergency Department or undergoing a primary care check-up. Haemoglobin A1c was measured from leftover whole blood samples using the enzymatic method on either Alinity c or Architect c-series analyser (Abbott Laboratories, Chicago, USA). Haemoglobin A1c between 39-47 mmol/mol was classified as prediabetes, while ≥ 48 mmol/mol as undiagnosed DM. RESULTS: After exclusion of 435 patients with known DM, the final cohort included 5092 patients (median age 57; 56% males). A total of 882 (17.3%) patients had HbA1c values between 39 and 47 mmol/mol. There were 214 (4.2%) patients with HbA1c ≥ 48 mmol/mol. Prediabetes prevalence ranged from 14.2% to 20.5%, while undiagnosed DM from 3.3% to 7.3%, with statistically significant differences among settings (P < 0.001). Age-stratified analysis showed that prediabetes and undiagnosed DM prevalence increase with age (P < 0.001), being 25.4% and 5.8%, respectively, in patients aged 60 to 70 years. CONCLUSION: Underlying impairment of glucose metabolism was identified in about one in five adults, with significant number of patients with already overt DM. These results should serve as a starting point for further steps directed towards promotion of preventive measures for DM in Croatia.

A prospective observational study of the relationship of critical illness associated hyperglycaemia in medical ICU patients and subsequent development of type 2 diabetes.

INTRODUCTION: Critical illness is commonly complicated by hyperglycaemia caused by mediators of stress and inflammation. Severity of disease is the main risk factor for development of hyperglycaemia, but not all severely ill develop hyperglycemia and some do even in mild disease. We hypothesised that acute disease only exposes a latent disturbance of glucose metabolism which puts those patients at higher risk for developing diabetes. METHODS: Medical patients with no history of impaired glucose metabolism or other endocrine disorder admitted to an intensive care unit between July 1998 and June 2004 were considered for inclusion. Glucose was measured at least two times a day, and patients were divided into the hyperglycaemia group (glucose ≥7.8 mmol/l) and normoglycaemia group. An oral glucose tolerance test was performed within six weeks after discharge to disclose patients with unknown diabetes or pre-diabetes who were excluded. Patients treated with corticosteroids and those terminally ill were also excluded from the follow-up which lasted for a minimum of five years with annual oral glucose tolerance tests. RESULTS: A five-year follow-up was completed for 398 patients in the normoglycaemia group, of which 14 (3.5%) developed type 2 diabetes. In the hyperglycaemia group 193 patients finished follow-up and 33 (17.1%) developed type 2 diabetes. The relative risk for type 2 diabetes during five years after the acute illness was 5.6 (95% confidence interval (CI) 3.1 to 10.2). CONCLUSIONS: Patients with hyperglycaemia during acute illness who are not diagnosed with diabetes before or during the hospitalization should be considered a population at increased risk for developing diabetes. They should, therefore, be followed-up, in order to be timely diagnosed and treated.

Obese patients in medical intensive care unit: influence of counseling on weight loss and cardiovascular parameters.

A case series of 12 obese patients admitted to medical intensive care unit (ICU) due to life-threatening diseases and the influence of weight loss on cardiovascular parameters is presented. We assessed body weight, body mass index (BMI), blood pressure, pulse, and laboratory values on admission. At discharge from ICU patients were counseled on how to lose weight. They were examined one and six months later. Statistically significant (p < 0.05) decrease of body weight (median at the beginning of a treatment 134 kg, after six months 127.5 kg), BMI (median 41.5 kg/m2 at the beginning of a treatment; 38.9 kg/m2 after six months), systolic blood pressure (medians 145 mmHg and 130 mmHg), diastolic blood pressure (medians 95 mmHg and 85 mmHg) and pulse (medians 104 beats per minute, 78 beats per minute) was found. The reduction of the waist circumference was not significant. One patient died due to severe acute pancreatitis. Patients reported feeling much better after losing weight.

[Possibilities of prevention of nephrotoxicity in intensive care unit]. / Mogucnosti prevencije nefrotoksicnog ucinka u intenzivnoj jedinici.

Different nephrotoxic drugs (antibiotic, antifungal, chemotherapeutic, NSAR, cyclosporine) as well as radio contrasts are mainly responsible for impairment of the renal funciton. Some patient populations, like diabetics, older patients and dehydrated patients are in additional risk for kidney failure. The challenge for protection of kidney function is complex, but it is possible in different clinical conditions to influence nephrotoxicity. The prevention of acute renal failure is very important, because this complication is according to many publications a significant risk for mortality. The administration of N acetilcystein, bicarbonate infusion, magnesium infusion, calcium channel blockers are promising strategies. It is very important to stress adequate rehydratation of every single patient, but some patient populations like diabetics, older patients and dehydrated patients are in special risk for nefrotoxicity. Because today radio contrast media are also important reason for nephrotoxicity, the use of low and iso-osmolar contrasts provides additional possibilities for prevention of potential nephrotoxicity.

[Hypothermia after cardiac arrest--underused method that saves lives]. / Hipotermija nakon kardijalnog aresta--premalo upotrebljavana metoda koja spasava zivote.

Cardiorespiratory arrest causes ischemia and lesion of all organ systems, but the central nervous system is the most vulnerable. It is known that only few minutes of hypoperfusion and ischemia can cause irreversible damage to the brain which is the major frustration of reanimatology. Results of clinical trials suggest positive effects of hypothermia on survival and neurological recovery which led to including this method to Guidelines for resuscitation as a recommended standard method in post-resuscitation period for patients who have not regained consciousness. Methods for induction and maintenance of hypothermia are numerous and various, basically divided into invasive and non-invasive, each with its own advantages and disadvantages which are described in this paper. Despite recognised positive effects of mild therapeutic hypothermia after resuscitation from cardiac arrest, the method is not fully implemented as a standard method in post-resuscitation period.

HbA1c is outcome predictor in diabetic patients with sepsis.

We have investigated predictive value of HbA1c for hospital mortality and length of stay (LOS) in patients with type 2 diabetes admitted because of sepsis. A prospective observational study was implemented in a university hospital, 286 patients with type 2 diabetes admitted with sepsis were included. Leukocyte count, CRP, admission plasma glucose, APACHE II and SOFA score were noted at admission, HbA1c was measured on the first day following admission. Hospital mortality and hospital length of stay (LOS) were the outcome measures. Admission HbA1c was significantly lower in surviving patients than in non-survivors (median 8.2% versus 9.75%, respectively; P<0.001). There was a significant correlation between admission HbA1c and hospital LOS of surviving patients (r=0.29; P<0.001). Logistic regression showed that HbA1c is an independent predictor of hospital mortality (odds ratio 1.36), together with female sex (OR 2.24), APACHE II score (OR 1.08) and SOFA score (OR 1.28). Multiple regression showed that HbA1c and APACHE II score are independently related to hospital LOS. According to our results, HbA1c is an independent predictive factor for hospital mortality and hospital LOS of diabetic patients with sepsis.

[Overall and therapy specific mortality of patients hospitalized for acute myocardial infarction in Zagreb University Hospital Centre]. / Razlike u mortalitetu bolesnika hospitaliziranih zbog akutnog infarkta miokarda u KBC-u Zagreb ovisno o nacinu lijecenja.

In Coronary Care Unit and Intensive Care Unit, Zagreb University Hospital Centre, 403 patients were hospitalized for acute myocardial infarction (AMI) in the year 2005. Majority (288) patients presented with ST-segment elevation myocardial infarction (STEMI) and 115 patients with acute myocardial infarction without ST-segment elevation (NSTEMI). In-hospital mortality of STEMI patients was 13.9% vs. 9.6% of NSTEMI group of patients. In STEMI group 202 patients underwent urgent percutaneous coronary intervention (PCI), and 86 patients in STEMI group received conservative therapy. In-hospital mortality of patients treated with primary PCI was 5.9% vs. 29.1% in the group treated conservatively. The main reason for conservative therapy was late presentation to the hospital (> 12 hours from the beginning of symptoms). Only 11 high-risk patients in NSTEMI group underwent primary PCI and 99 NSTEMI patients received conservative therapy (in-hospital mortality 10.1%). We performed totally 218 urgent PCI interventions in both groups with low in-hospital mortality of 6.1% in comparison with high mortality in conservatively treated groups of patients. The mortality in PCI group was strongly connected with unsuccessful intervention and late reperfusion. Total ischemic time (6 hours and 5 minutes), and time from hospital presentation to reperfusion -"door to balloon time" (1 hour and 25 minutes) are longer that in similar patients series, and need to be improved. Transportation time (1 hour and 40 minutes) is acceptable. In conclusion, high percentage (70.1%) of STEMI and NSTEMI patients underwent primary PCI in our institution with low perioperative mortality. This group of patients had superior in-hospital mortality when compared with conservatively treated group of patients. There is still unacceptably high percentage of patients with AMI, who came into the hospital too late for any reperfusion therapy.

Sepsis syndrome in Croatian intensive care units: piloting a national comparative clinical database.

AIM: To assess the incidence of sepsis in selected intensive care units (ICUs) in Croatia, isolates from blood cultures, and sepsis outcomes, and to compare the results with those from other European countries. METHODS: In the pilot phase of the national comparative clinical database project, we included 24 ICUs--general, specialized, neonatal, pediatric, and adult--18 from university hospitals, 3 from county hospitals, and 4 from city hospitals. By retrospective chart review, trained data collectors abstracted the data on the case mix, management strategies, and outcomes in patients consecutively admitted to ICUs. Central validation for incomplete, illogical, or inconsistent values is regularly performed to improve accurateness. RESULTS: Of 5293 patients treated in 24 ICUs from November 1, 2004, to October 31, 2005, 456 (8.6%) were treated for sepsis syndrome or severe sepsis. The most common isolates from positive blood cultures were Esherichia coli (11.6%), Pseudomonas species (9.9%), and methicillin-resistant Staphylococcus aureus (9.3%). With the mean Acute Physiology and Chronic Health Evaluation (APACHE) II score of 10.0 and Sequential Organ Failure Assessment (SOFA) score of 2.4, the overall mortality for sepsis syndrome, severe sepsis, and septic shock was 29%, 35%, and 34%, respectively. When compared to university hospitals and county hospitals, city hospitals with the smallest gravitating population had significantly lower APACHE II and SOFA scores, but significantly higher mortality. CONCLUSIONS: Overall mortality of patients with sepsis syndrome in Croatian ICUs was high, but outcomes of their treatment were comparable with those in other European countries. Better education in triage and treatment strategies is needed, including better implementation of Surviving Sepsis Campaign guidelines.

Hemophagocytic syndrome--should we consider it more often?

Hemophagocytic syndrome (HPS) is a rare condition characterized by overactive histiocytes, hepatosplenomegaly, fever and cytopenia, with two major types: familial, autosomal recessive genetic disease and acquired that can occur during systemic infections, immunodeficiency or malignancy. Inappropriate activation of macrophages by cytokines is the major mechanism of the disease. We report a case of an adult patient with HPS. After thorough clinical investigation, we have not been able to establish the underlying disease, and corticosteroids therapy was initiated empirically. After 8 months follow-up the patient is well with normal laboratory findings.