Comparison of articular and auricular cartilage as a cell source for the autologous chondrocyte implantation

Articular (medial femoral condyle) and auricular cartilage (anithelix) was compared as a cell source for the autologous joint repair. Cells isolated from five human cadaveric donors were cultured parallel in the monolayer cultures and in the 3D alginate hydrogel constructs for 1 week. Cell morphology was controlled by the fluorescent microscopy and gene expressions of type I collagen (COL1), type II collagen (COL2), aggrecan (AGR), versican (VER), and elastin (ELS) were analyzed by the real‐time polymerase chain reaction. COL1 and ELS, predominant in the phenotype of auricular biopsy, were statistically lower in the articular biopsies. Even though COL2 and AGR decreased in monolayers of both cell sources, the dedifferentiation process affected auricular cells intensely. Cells embedded in the alginate hydrogel directly after the isolation did not exhibit the dedifferentiated phenotype. Additionally, COL1, COL2, AGR, and VER were comparable between the two sources. ELS however, remained higher in the auricular cells regardless of the culture type. The study indicates that auricular chondrocytes cultured in a 3D environment immediately after the isolation have a neo‐cartilage potential for the articular surface reconstruction. © 2008 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 27: 943–948, 2009     

Characteristics of heart rate variability in war veterans with post-traumatic stress disorder after myocardial infarction

OBJECTIVE: The goal of the study was to evaluate differences in heart rate variability (HRV) among post-myocardial infarction (MI) patients, depending on their participation in the Croatian war and on established diagnoses of post-traumatic stress disorder (PTSD). METHODS: The study included 34 male war veterans with diagnosed PTSD who had suffered a first MI and 34 age-matched post-MI patients without PTSD. Cardiac autonomic balance was evaluated through HRV analysis. RESULTS: There were no differences in the mean R-R interval or overall HRV between the analyzed groups. Post-MI patients with PTSD had lower values for the square root of the mean of squared successive differences in R-R intervals (p = 0.02), the percentage of R-R intervals that were > or =50 milliseconds different from the previous interval (p = 0.03), and the high-frequency component (p = 0.03) but had higher values for the low-frequency component (p = 0.01) and the low-frequency/high-frequency ratio (p = 0.02), compared with post-MI patients without PTSD. CONCLUSION: Post-MI patients with PTSD have higher sympathetic and lower parasympathetic heart rate modulation activity, compared with patients with MI and no PTSD.     

Irinotecan toxicity to human blood cells in vitro: relationship between various biomarkers

Toxic effects of the antineoplastic drug irinotecan on human blood cells at concentrations of 9.0 microg/ml and 4.6 microg/ml were evaluated in vitro. Using the alkaline and neutral comet assay significantly increased levels of primary DNA damage in lymphocytes were detected. The induction of apoptosis/necrosis, as determined by a fluorescent assay, was also notably increased. Cytogenetic outcomes of the treatment were assessed by the analysis of structural chromosome aberrations and fluorescence in situ hybridization. A significantly higher incidence of chromatid breaks and complex quadriradials was observed. Painted chromosomes 1, 2 and 4 were equally involved in translocations, but only the chromosome 1 was involved in the formation of quadriradials. Sister chromatid exchange analysis was performed in parallel with the analysis of lymphocyte proliferation kinetics. The higher concentration of irinotecan caused almost seven-time increase, while the lower one caused a five-time increase of the basal sister chromatid exchange frequency, accompanied with significant lowering of the lymphocyte proliferation index. Using the cytokinesis-block micronucleus assay, a dose-dependent increase in micronucleus frequency along with the formation of nuclear buds and nucleoplasmic bridges was noticed. Inhibitory effects of irinotecan on enzyme acetylcholinesterase (AChE) were studied in erythrocytes. An IC(50) value of 5.0 x 10(-7) was established. Irinotecan was found to be strong inhibitor of the acetylcholine hydrolysis and to cause a continuous decrease of catalytic activity of AChE. The results obtained on a single donor may contribute to the understanding of irinotecan toxicity, but further in vitro and in vivo studies are essential in order to clarify remaining issues, especially on possible inter-individual variability in genotoxic responses to the drug.     

Antiepileptic, behavioral, and antidepressant effects of adjuvant lamotrigine therapy in drug-resistant epilepsy

AIM: To evaluate the behavioral effects of lamotrigine as add-on therapy in treatment-resistant epilepsy. METHODS: An open, prospective, long-term study of lamotrigine as adjuvant therapy was performed in 56 patients with drug-resistant epilepsy (female/male ratio 35/21, age range 16-51 years). All the patients kept seizure diaries, and electroencephalograms were recorded at baseline and during 24 months of the treatment. Quality of life questionnaire, Hamilton depression scale (HMD), Beck depression scale (BDI), and Hamilton anxiety scale (HMA) were used before and during lamotrigine therapy. Comparative assessments were made in an age- and sex-matched control group treated with other antiepileptic drugs. RESULTS: Overall, seizure control was improved in 55.3% of the patients, remained unchanged in 39.3%, and deteriorated in 5.4%. Improvement in some quality of life measures occurred in 50% of the patients. The HMD subscales and BDI scale showed significant improvement in lamotrigine treated patients compared to the control group (ANOVA, p < 0.01). Negative behavioral effects occurred in 10.7% of the patients. CONCLUSION: Lamotrigine demonstrated significant antiepileptic long-term efficacy, and its positive effects on the mood and quality of life, which surpassed the negative behavioral effects, and contributed highly to the favorable treatment outcome.     

In Vivo Evaluation of Cholinesterase Activity,Oxidative Stress Markers,Cyto‐ and Genotoxicity of K048 Oxime – a Promising Antidote against Organophosphate Poisoning

K048 is a member of K‐oximes, a new oxime class that has recently been confirmed effective against poisoning by the nerve agent tabun and several pesticides. The toxicity profile of the K048 oxime has not been fully characterized and its optimal therapeutic dose has not yet been established. Earlier studies report excellent results with K048 in reactivating tabun‐phosphorylated AChE and in the therapy of tabun‐poisoned mice. It possesses a low acute toxicity and exerts an acceptable toxicity profile on isolated human peripheral blood lymphocytes in vitro. Intraperitoneal administration of K048 in rats resulted in an LD₅₀ of 238.3 mg/kg. In this in vivo study, we investigated cholinesterase (ChE) activity and oxidative stress marker levels (lipid peroxidation and superoxide dismutase activity) in the plasma of exposed rats after administering the compound at 25% of its LD₅₀. Lymphocyte viability was evaluated using an acridine orange/ethidium bromide in situ fluorescent assay. The levels of primary DNA damage in rat white blood cells were measured using the alkaline comet assay. The compound applied at 25% of its LD₅₀ did not significantly affect ChE activity and lipid peroxidation and did not cause significant changes in the SOD activity in plasma. The cytotoxicity profile of K048 in the tested dose was also acceptable, and it did not possess significant DNA‐damaging potential. The obtained results are promising for further evaluations of the K048 oxime, which should include tests on a broader concentration range and longer incubation times.     

Trace Elements in Tissues of Wild Carnivores and Omnivores in Croatia

The differences in metal exposure (As, Cd, Cu, Pb and Hg) in the muscle, liver and kidney tissues of brown bears (Ursus arctos), grey wolfs (Canis lupus), Eurasian lynxs (Lynx lynx), Eurasian badgers (Meles meles) and pine martens (Martes martes) from Croatia were observed. The highest mean Cd levels were found in kidney and liver of Eurasian badger (3.05 and 0.537 mg/kg). The highest Cu concentrations (mg/kg) measured in liver tissue were obtained in order: Eurasian badger (15.2) > brown bear (12.1) > pine marten (10.3) > Eurasian lynx (8.43) > grey wolf (6.44). Result presented that Eurasian badger accumulated the highest levels of elements: As, Cu and Pb in muscle; As, Cd, Cu and Pb in liver; Cd and Pb in kidney. Kidney of pine marten accumulated the highest concentrations of As, Cu and Hg. Omnivorous species observed present an important bioindicator for the accumulation of toxic elements indicating an enhanced vulnerability for response to ecological changes in forested terrain. Generally, element concentrations found in five species observed were lower in comparison to levels reported in previous studies and below levels related to toxicosis in mammals.     

Real-life experience with remdesivir for treatment of hospitalized coronavirus disease 2019 patients: matched case-control study from a large tertiary hospital registry

AimTo evaluate the association of remdesivir use and the survival of hospitalized patients with coronavirus disease 2019 (COVID-19).MethodsWe retrospectively reviewed the medical records of 5959 COVID-19 patients admitted to our tertiary-level hospital from March 2020 to June 2021. A total of 876 remdesivir-treated patients were matched with 876 control patients in terms of age, sex, Charlson comorbidity index (CCI), WHO-defined COVID-19 severity on admission, and oxygen requirement at the time of remdesivir use.ResultsAmong 1752 COVID-19 patients (median age 66 years, 61.8% men), 1405 (80.2%) had severe and 311 (17.8%) had critically severe COVID-19 on admission. Remdesivir was given at a median of one day after hospital admission and at a median of eight days from the onset of symptoms. Overall, 645 (73.6%) patients received remdesivir before high-flow oxygen therapy (HFOT) or mechanical ventilation (MV), 198 (22.6%) after HFOT institution, and 83 (9.5%) after MV institution. Remdesivir use was associated with improved survival in the entire cohort (hazard ratio 0.79, P = 0.006). Survival benefit was evident among patients receiving remdesivir during low-flow oxygen requirement (hazard ratio 0.61, P < 0.001) but not among patients who received it after starting HFOT (P = 0.499) or MV (P = 0.380).ConclusionRemdesivir, if given during low-flow oxygen therapy, might be associated with survival benefit in hospitalized COVID-19 patients.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection affects multiple organ systems and, in a substantial proportion of patients, presents itself through severe or critical symptoms (1,2). Patients developing respiratory insufficiency require hospital admission and often critical or intensive care. Vaccination reduced the number of patients with severe disease presentation who require hospital admission, thus leading to better outcomes in those who developed severe disease despite having received the vaccine (3). However, vaccine hesitancy (4) and waning effects of vaccination (5) remain important problems for long-term coronavirus disease 2019 (COVID-19) control.Remdesivir is the first antiviral drug to be approved for the treatment of severe or critical COVID-19 based on shortened time to recovery (6), but it demonstrated no clear survival benefit in the randomized controlled setting (7). Reports based on real-life cohorts of COVID-19 patients suggest survival benefit of remdesivir in patients with less severe disease (8-11). However, the results are inconsistent (12,13), and the reports are conflicting regarding certain aspects of remdesivir use, such as the length of hospital stay (13). Due to uncertainties regarding the efficacy of remdesivir among real-life patients burdened with comorbidities, we evaluated the association of remdesivir use with survival in a large real-life cohort of COVID-19 patients treated in our tertiary level institution.     

Incidence and risk factors for venous and arterial thromboses in hospitalized patients with coronavirus disease 2019: data on 4014 patients from a tertiary center registry

AimTo evaluate the burden and predictors of thromboembolic complications in a large real-life cohort of hospitalized patients with established coronavirus disease 2019 (COVID-19).MethodsWe retrospectively reviewed the records of 4014 consecutive adult patients admitted to a tertiary-level institution because of COVID-19 from March 2020 to March 2021 for the presence of venous and arterial thrombotic events.ResultsVenous-thromboembolic (VTE) events were present in 5.3% and arterial thrombotic events in 5.8% patients. The majority of arterial thromboses occurred before or on the day of admission, while the majority of VTE events occurred during hospitalization. The majority of both types of events occurred before intensive care unit (ICU) admission, although both types of events were associated with a higher need for ICU use and prolonged immobilization. In multivariate logistic regression, VTE events were independently associated with metastatic malignancy, known thrombophilia, lower mean corpuscular hemoglobin concentration, higher D-dimer, lower lactate dehydrogenase, longer duration of disease on admission, bilateral pneumonia, longer duration of hospitalization, and immobilization for at least one day. Arterial thromboses were independently associated with less severe COVID-19, higher Charlson comorbidity index, coronary artery disease, peripheral artery disease, history of cerebrovascular insult, aspirin use, lower C reactive protein, better functional status on admission, ICU use, immobilization for at least one day, absence of hyperlipoproteinemia, and absence of metastatic malignancy.ConclusionAmong hospitalized COVID-19 patients, venous and arterial thromboses differ in timing of presentation, association with COVID-19 severity, and other clinical characteristics.

An increasing pool of evidence accumulating since the early days of the coronavirus disease 2019 (COVID-19) pandemic shows that severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection induces pro-thrombotic state. Although the disease presents dominantly with respiratory symptoms, resulting in acute respiratory distress syndrome in a subset of patients (1-4), high frequencies of venous and arterial thromboses were observed (5). SARS-CoV-2 endothelial tropism and damage to the vasculature of the lungs, heart, extremities, and brain have been recognized as the key part of disease pathophysiology (6-9). In addition, coagulopathy due to upregulation of inflammatory prothrombotic proteins, platelet activation, and immobilization of patients due to functional deterioration or connection to the oxygen source further favor the development of thrombotic incidents. Vascular complications occur mostly in severe cases and are often associated with multiorgan failure and higher mortality. In some patients, they can also be the dominant clinical presentation. The most frequently noted thrombotic events are pulmonary embolism (PE) and deep vein thrombosis (DVT) (10), followed by stroke, acute limb ischemia, and acute coronary syndromes (11,12), developing despite the use of pharmacologic thromboprophylaxis in hospitalized COVID-19 patients.Vascular endothelial injury is not COVID-19 specific, as similar response was described in other acute infectious diseases (13,14). However, the large scale of the COVID-19 pandemic and associated strain on the health care system result in a substantial number of patients urgently presenting to the hospital or being under an increased risk of development of thromboembolic complications. Due to importance of this issue and scarcity of regional data that may help guide the medical care of COVID-19 patients, studies on this issue are highly needed. Thus, the aim of this study was to evaluate the burden of thromboembolic complications and their predictors in a large real-life cohort of hospitalized patients with established COVID-19.     

The variable number of tandem repeat polymorphism of platelet glycoprotein Ibalpha and risk of coronary heart disease

Glycoprotein (GP) Ib-IX-V complex plays an important role in formation of platelet-fibrin clot at the area of damaged vessel wall. One polymorphism of GP Ibalpha, the main component of GP Ib-IX-V complex, is due to variable numbers of tandem repeats (VNTRs) in the macroglycopeptide region of this molecule. We studied the association between the presence of different VNTR alleles of GP Ibalpha and the frequency of coronary heart disease (CHD) among individuals recruited to a large community-based case-cohort study (Atherosclerosis Risk in Communities [ARIC] study). We found that the distribution of VNTR alleles of GP Ibalpha is different among whites and African Americans. The B allele (with 3 repeats) of GP Ibalpha is relatively more common among African Americans compared with whites. In African Americans, the CC genotype (homozygous with 2 repeats) is associated with a lower risk of CHD events than all other genotypes.