Influence of hemolysis on clinical chemistry parameters determined with Beckman Coulter tests – detection of clinically significant interference

The aim of this study was to examine the influence of hemolysis on 25 clinical chemistry parameters and to compare the resulting bias with clinically significant differences and the manufacturer’s specifications. Using freeze-thawing of the treated blood aliquot of each subject (N?=?17), four hemolysis levels were prepared with hemolysis index (HI) and hemoglobin concentration as follows: (+)=0.5–0.99?g/L, (2+)=1–1.99?g/L, (3+)=2–2.99?g/L and (4+)=3–4.99?g/L. All analytes were tested on the Beckman Coulter AU480 analyzer using proprietary reagents. It was considered that the interference was detected if the 95% confidence interval for mean differences (%) between hemolyzed and non-hemolyzed samples did not include zero. Clinically significant interference was judged against reference change value (RCV). Hemolysis interference was detected for: alpha-amylase, alkaline phosphatase (ALP), aspartate aminotransferase (AST), total and conjugated bilirubin, creatine kinase (CK), CK-MB, ?-glutamyltransferase (GGT), iron, lactate dehydrogenase (LD), magnesium, potassium, total protein and uric acid at HI=(1+); alanine aminotransferase (ALT) and phosphate at HI=(2+); urea at HI=(3+); albumin and cholinesterase at HI=(4+). Even at the greatest hemolysis degree, HI=(4+), no interference was detected for calcium, chloride, creatinine, C-reactive protein (CRP), glucose and sodium. Clinically significant difference was exceeded for LD at HI=(1+); CK-MB at HI=(2+); AST and potassium at HI=(3+); total bilirubin at HI=(4+). The presented results did not support the manufacturer’s claim for CK and GGT. Establishing HI thresholds for reporting or suppressing test results is the responsibility of each laboratory, taking into account the manufacturer’s data, but also its own investigations.     

Pentadecapeptide BPC 157 resolves suprahepatic occlusion of the inferior caval vein,Budd-Chiari syndrome model in rats

BACKGROUND Recently, as a possible therapy resolving solution, pentadecapeptide BPC 157 therapy, has been used in alleviating various vascular occlusion disturbances.BPC 157 was previously reviewed as novel mediator of Robert cytoprotection and endothelium protection in the stomach, and gut-brain axis, beneficial therapy in gastrointestinal tract, with particular reference to vascular recruitment, ulcerative colitis and tumor cachexia, and other tissues healing. Here we raised new hypothesis about BPC 157 therapy in the Budd-Chiari syndrome in rats, rapid bypassing of the suprahepatic inferior caval vein occlusion, and rats recovery with the active and effective pharmacotherapy treatment.AIM To investigate Budd-Chiari syndrome model(inferior caval vein suprahepatic occlusion) resolution, since BPC 157 resolves various rat vascular occlusion.METHODS We assessed the activated bypassing pathways between the inferior and superior caval veins and portocaval shunt, counteracted caval/portal hypertension, aortal hypotension, venous/arterial thrombosis, electrocardiogram disturbances, liver and gastrointestinal lesions(i.e., stomach and duodenum hemorrhages, in particular, congestion). Rats with suprahepatic occlusion of the inferior vena cava by ligation were medicated at 1 min, 15 min, 24 h, or 48 h post-ligation.Medication consisted of 10 μg/kg BPC 157, 10 ng BPC 157 or 5 m L/kg saline,administered once as an abdominal bath or intragastric application. Gross and microscopic observations were made, in addition to assessments of electrical activity of the heart(electrocardiogram), portal and caval hypertension, aortal hypotension, thrombosis, hepatomegaly, splenomegaly and venography.Furthermore, levels of nitric oxide, malondialdehyde in the liver and serum enzymes were determined.RESULTS BPC 157 counteracted increased P wave amplitude, tachycardia and ST-elevation,i.e., right heart failure from acute thrombotic coronary occlusion. The bypassing pathway of the inferior vena cava-azygos(hemiazygos) vein-superior vena cava and portocaval shunt occurred rapidly. Even with severe caval portal hypertension, BPC 157 antagonized portal and caval hypertension and aortal hypotension, and also reduced refractory ascites. Thrombosis of portal vein tributaries, inferior vena cava, and hepatic and coronary arteries was attenuated.In addition, there was reduced pathology of the lungs(severe capillary congestion) and liver(dilated central veins and terminal portal venules),decreased intestine hemorrhagic lesions(substantial capillary congestion,submucosal edema and architecture loss), and increased liver and spleen weight.During the period of ligation, nitric oxide-and malondialdehyde-levels in the liver remained within normal healthy values, and increases in serum enzymes were markedly reduced.CONCLUSION BPC 157 counteracts Budd Chiari syndrome in rats.     

Assessment of cytotoxic and genotoxic effects of conventional and whitening kinds of toothpaste on oral mucosa cells

Objective: This study aimed to evaluate possible DNA damages to oral epithelial cells exposed to whitening kinds of toothpaste considering the effect of conventional non-whitening toothpaste.

Materials and methods: Sixty volunteers were assigned into three experimental groups, each of them using a different regular toothpaste for the initial 2 months, followed by the use of whitening kind of toothpaste of the same brand for next 2 months. The oral epithelial cells were sampled prior and 30, 60, 90 and 120 days after the beginning of the use of tested kinds of toothpaste. Chromosomal damages were analyzed by micronucleus assay.

Results: For just one kind of tested whitening toothpaste was observed the significant increase in the number of micronucleated cells after 60 days of use compared values obtained 60 days of usage of conventional non-whitening toothpaste (6.35?±?3.67 and 2.8?±?1.91; p?Conclusions: Based on the results, it can be concluded that the use of certain whitening kinds of toothpaste may cause a limited biologically insignificant genotoxic effect on buccal epithelial cells.     

FlnA-null megakaryocytes prematurely release large and fragile platelets that circulate poorly

Filamin A (FlnA) is a large cytoplasmic protein that crosslinks actin filaments and anchors membrane receptors and signaling intermediates. FlnA(loxP) PF4-Cre mice that lack FlnA in the megakaryocyte (MK) lineage have a severe macrothrombocytopenia because of accelerated platelet clearance. Macrophage ablation by injection of clodronate-encapsulated liposomes increases blood platelet counts in FlnA(loxP) PF4-Cre mice and reveals the desintegration of FlnA-null platelets into microvesicles, a process that occurs spontaneously during storage. FlnA(loxP) PF4-Cre bone marrows and spleens have a 2.5- to 5-fold increase in MK numbers, indicating increased thrombopoiesis in vivo. Analysis of platelet production in vitro reveals that FlnA-null MKs prematurely convert their cytoplasm into large CD61(+) platelet-sized particles, reminiscent of the large platelets observed in vivo. FlnA stabilizes the platelet von Willebrand factor receptor, as surface expression of von Willebrand factor receptor components is normal on FlnA-null MKs but decreased on FlnA-null platelets. Further, FlnA-null platelets contain multiple GPIbα degradation products and have increased expression of the ADAM17 and MMP9 metalloproteinases. Together, the findings indicate that FlnA-null MKs prematurely release large and fragile platelets that are removed rapidly from the circulation by macrophages.     

A Slovenian Model of Comprehensive Care for Patients with Difficulties Accessing Healthcare: A Step Towards Health Equity

ABSTRACT

Patients that cannot come to their family medicine practice (i.e. who have difficulties with access) do not receive the same preventive screening activities and management of their chronic diseases as those who can. Community nurses who provide healthcare to patients in their homes were trained in additional competencies, including screening for risk factors for selected diseases and the management of patients with selected chronic diseases. The presented model enables equal management of all registered patients, regardless of accessibility. It also fosters exchange of information within the team members and thus improves the quality of the team management of patients.     

Fever of unknown origin: large vessel vasculitis diagnosed by PET/CT

PET/CT is starting to play an important role in evaluating fever of unknown origin (FUO), due to its ability to localize and delineate areas of high metabolic activity, such as neoplastic proliferation and inflammation, including vasculitis. We present a case of giant cell arteritis (GCA) in a 72-year-old female patient admitted to our department with a 4-month history of FUO, weight loss and fatigue, without specific symptoms or signs. Laboratory investigations suggested acute phase response, with a pronounced erythrocyte sedimentation rate, high CRP level and microcytic anemia. A thorough diagnostic evaluation was performed to exclude an unknown primary tumor, which was initially suspected due to a positive family history of cancer. Surprisingly, PET/CT revealed large vessel vasculitis affecting the ascending, descending and abdominal aorta, as well as subclavian, proximal brachial and carotid arteries bilaterally. Biopsy of the superficial temporal artery confirmed the diagnosis of GCA. Treatment with methylprednisolone and azathioprine led to resolution of clinical symptoms and normalization of laboratory parameters. In addition to the use of PET/CT in the evaluation of FUO, its value as a method complementary to temporal artery biopsy is also discussed.     

The laparoscopic liver resections—an initial experience and the literature review

The laparoscopic liver resection (LLR) represents a new pathway in hepatic surgery. Several studies have reported its application in both malignant and benign liver diseases. The most common liver resections performed laparoscopically are wedge, segmental resections and metastasectomy; although in large centers the laparoscopic right and left hepatectomies have begun to perform more frequently. We report the initial experience in LLRs at our department including a case of the first laparoscopic left lateral liver bisegmentectomy performed in patient with follicular nodular hyperplasia and the 15 cases of wedge laparoscopic resections of echinococcic liver cysts. According to literature the mortality rate in LLRs is up to 0.3% and morbidity rate up to 10.5%. The most common cause of the death is liver failure, while the most frequent complication is the bile leakage. Advantages for patients include smaller incisions, less blood loss, and shorter lengths of hospital stay. The LLRs in experienced hands were shown to be safe with acceptable morbidity and mortality for both minor and major hepatic resections in benign and malignant diseases.     

Epidemiological characteristics,baseline clinical features,and outcomes of critically ill patients treated in a coronavirus disease 2019 tertiary center in continental Croatia

AimTo describe epidemiological characteristics and baseline clinical features, laboratory findings at intensive care unit (ICU) admission, and survival rates of critically ill coronavirus disease 2019 (COVID-19) patients treated at a tertiary institution specialized for COVID-19 patients.MethodsThis retrospective study recruited 692 patients (67.1% men). Baseline demographic data, major comorbidities, anthropometric measurements, clinical features, and laboratory findings at admission were compared between survivors and non-survivors.ResultsThe median age was 72 (64-78) years. The median body mass index was 29.1 kg/m². The most relevant comorbidities were diabetes mellitus (32.6%), arterial hypertension (71.2%), congestive heart failure (19.1%), chronic kidney disease (12.6%), and hematological disorders (10.3%). The median number of comorbidities was 3 and median Charlson Comorbidity Index (CCI) was 5. A total of 61.8% patients received high-flow nasal oxygen therapy (HFNO) and 80.5% received mechanical ventilation (MV). Median duration of HFNO was 3, and that of MV was 7 days. ICU mortality rate was 72.7%. Survivors had significantly lower age, number of comorbidities, CCI, sequential organ failure assessment score, serum ferritin, C-reactive protein, D-dimer, and procalcitonin, interleukin-6, lactate, white blood cell, and neutrophil counts. They also had higher lymphocyte counts, PaO₂/FiO₂ ratio, and glomerular filtration rate at admission. Length of ICU stay was 9 days. The median survival was 11 days for mechanically ventilated patients, and 24 days for patients who were not mechanically ventilated.ConclusionThe parameters that differentiate survivors from non-survivors are in agreement with published data. Further multivariate analyses are warranted to identify individual mortality risk factors.

The first case of coronavirus disease 2019 (COVID-19) in Croatia was confirmed on February 25, 2020 (1). Very soon, on March 11, the World Health Organization (WHO) declared a COVID-19 pandemic (2). As of February 25, 2021, there were more than 240 000 confirmed cases and 5489 deaths in Croatia.As a part of the national strategy against COVID-19 pandemic, the Ministry of Health and Civil Protection Headquarters decided that University Hospital Dubrava (UH) is to be repurposed into a Primary Respiratory Center for patients with confirmed COVID-19 infection. The intensive center of primary respiratory intensive center (PRIC-IC) is a subunit of UH Dubrava reserved for the treatment of patients with severe symptoms of COVID-19 who require mechanical ventilation, vasoactive hemodynamic support, continuous renal replacement therapy, and other aspects of intensive care (3). UH Dubrava became a COVID-19 tertiary center treating a third of all COVID-19 positive ICU patients in the country.As the pandemic was surging through Europe, the number of critically ill COVID-19 patients in UH Dubrava continued to grow, and ICU capacities needed expansion. During winter months, six intensive care units in PRIC were operating at the same time: Three were run by intensivists from UH Dubrava and three by intensivists from other hospitals in Zagreb, including University Hospital Center Zagreb, University Hospital Center Sestre Milosrdnice, University Hospital Sveti Duh, University Hospital Merkur, and Children''s Hospital Zagreb. The outcomes of critically ill patients treated in PRIC-IC therefore represent the work of intensivists from all hospitals in Zagreb.Although scientific knowledge of COVID-19 increases daily, limited information is available regarding early identification of individuals who are at risk of developing severe symptoms. Previous studies reported certain demographic features and clinical characteristics of patients who were likely to develop severe symptoms of COVID-19 and thus require mechanical ventilation (4-7). Studies worldwide reported high mortality rates for patients requiring mechanical ventilation, ranging from 40% to 97% (4,8-10). Unfortunately, some of these reports were preliminary and included patients without a completed ICU stay. The aim of our cohort retrospective study is to describe the demographic characteristic, clinical features, laboratory values, and outcomes among critically ill COVID-19 patients treated in PRIC-IC, UH Dubrava.