Dose-dependent effects of acetaminophen and ibuprofen on mitochondrial respiration of human platelets.

Acetaminophen and ibuprofen are widely used over-the-counter medications to reduce fever, pain, and inflammation. Although both drugs are safe in therapeutic concentrations, self-medication is practiced by millions of aged patients with comorbidities that decrease drug metabolism and/or excretion, thus raising the risk of overdosage. Mitochondrial dysfunction has emerged as an important pathomechanism underlying the organ toxicity of both drugs. Assessment of mitochondrial oxygen consumption in peripheral blood cells is a novel research field Cu several applications, including characterization of drug toxicity. The present study, conducted in human platelets isolated from blood donor-derived buffy coat, was aimed at assessing the acute, concentration-dependent effects of each drug on mitochondrial respiration. Using the high-resolution respirometry technique, a concentration-dependent decrease of oxygen consumption in both intact and permeabilized platelets was found for either drug, mainly by inhibiting complex I-supported active respiration. Moreover, ibuprofen significantly decreased the maximal capacity of the electron transport system already from the lowest concentration. In conclusion, platelets from healthy donors represents a population of cells easily available, which can be routinely used in studies assessing mitochondrial drug toxicity. Whether these results can be recapitulated in patients treated with these medications is worth further investigation as potential peripheral biomarker of drug overdose.

Toxoplasma gondii Infection in Patients with Cardiovascular Diseases from Western Romania: A Case-Control Study.

(1) Background: Toxoplasma gondii infects approximately one third of the world's human population. The seroepidemiology of T. gondii in cardiovascular patients is poorly discussed in the existing literature. We aimed to evaluate, for the first time, the seroprevalence of T. gondii in cardiovascular patients from Western Romania. (2) Methods: Serologic testing to demonstrate the presence of T. gondii antibodies was conducted in 256 patients with cardiovascular diseases and 261 matched blood donors. (3) Results: The overall seroprevalence of T. gondii antibodies was 64.06% in patients with cardiovascular diseases and 52.88% in blood donors and tended to increase with age in both groups. The seroprevalence of T. gondii antibodies was significantly higher in cardiovascular male patients (69.94%) compared to male blood donors (55.69%) (p = 0.006). When compared to the control group, a significantly higher prevalence of T. gondii antibodies was found among patients with hypertension (82.35%; p = 0.01) and unstable angina (67.56%; p = 0.02). (4) Conclusions: This study brings new epidemiological information on the prevalence of T. gondii in Romanian cardiovascular patients. T. gondii seroprevalence was significantly higher in patients with hypertension and unstable angina, suggesting that individuals with these diagnoses may be more frequently infected with T. gondii. This study may be a valuable starting point for further research to better evaluate the impact of T. gondii exposure on patients with cardiovascular diseases.

Seroprevalence of Anti-Echinococcus granulosus Antibodies and Risk Factors for Infection in Blood Donors from Western Romania.

Cystic echinococcosis is a worldwide-distributed zoonotic parasitic disease. This cross-sectional study aimed to assess the seroprevalence and risk factors potentially associated with Echinococcus granulosus in healthy blood donors from Timis County, an endemic region in Western Romania. Serum samples were collected from 1347 Romanian blood donors. Serologic tests to determine the presence of anti-Echinococcus antibodies were performed using an anti-Echinococcus-ELISA immunoassay. Anti-Echinococcus antibodies were detected in 38 blood donors, indicating an overall seroprevalence of 2.8%. The seropositivity rate was 3.7% in females and 3.1% in blood donors residing in urban areas. The highest seropositivity was found in the age group of 31-40 years (3.6%). There were no significant differences between Echinococcus seropositivity and gender, area of residence, age, contact with dogs, or raising sheep. This serologic survey evaluated for the first time the presence of Echinococcus antibodies in healthy blood donors from Western Romania and the potential risk factors associated with echinococcosis. Our results suggest that this zoonotic infection might evolve asymptomatically in apparently healthy individuals. Further studies should be conducted in the general population to estimate the true extent of human echinococcosis and its risk factors.

Monoamine oxidase is a source of cardiac oxidative stress in obese rats: the beneficial role of metformin.

Diet-induced metabolic diseases, such as obesity, metabolic syndrome, and type 2 diabetes (T2DM) are the global threatening epidemics that share cardiovascular oxidative stress as common denominator. Monoamine oxidase (MAO) has recently emerged as a constant source of reactive oxygen species (ROS) in DM. Metformin, the first-line drug in T2DM, elicits cardiovascular protection via pleiotropic effects. The present study was aimed to assess the contribution of MAO to the early cardiac oxidative stress in a rat model of high-calorie junk food (HCJF) diet-induced obesity and prediabetes and whether metformin can alleviate it. After 6 months of HCJF, rats developed obesity and hyperglycemia. Hearts were isolated and used for the evaluation of MAO expression and ROS production. Experiments were performed in the presence vs absence of metformin (10 µM) and MAO-A and B inhibitors (clorgyline and selegiline, 10 µM), respectively. Both MAO isoforms were overexpressed and led to increased ROS generation in cardiac samples harvested from the obese animals. Acute treatment with metformin and MAO inhibitors was able to mitigate oxidative stress. More important, metformin downregulated MAO expression in the diseased samples. In conclusion, MAO contributes to oxidative stress in experimental obesity and can be targeted with metformin.

Seroepidemiology of Toxoplasma gondii Infection in Blood Donors from Western Romania.

Background: Toxoplasma gondii is estimated to infect 33% of blood donors worldwide, and seroprevalence varies widely between countries. We aimed to evaluate, for the first time, the seroprevalence and risk factors associated with T. gondii in blood donors from Western Romania. Methods: Serologic testing to demonstrate the presence of T. gondii antibodies was conducted in 1347 healthy blood donors. Risk factors for T. gondii infection were assessed through an epidemiological questionnaire. Results: The overall prevalence of T. gondii antibodies was 45.9%, with a significant age-associated increase (p < 0.001) from 32.6% in age group 18−25 years to 67.6% in age group 56−63 years. T. gondii seroprevalence decreased with increasing level of education, from 64.3% in individuals who graduated from elementary/middle school to 40.4% in those who graduated from University (p < 0.001). The multiple logistic regression analysis revealed that age, level of education and having pets (cats and/or dogs) were significantly associated with T. gondii infection. Conclusions: This study brings new and valuable data regarding the seroepidemiology of T. gondii infection in Romania. Our findings indicate a high prevalence of T. gondii antibodies in blood donors and may serve as a starting point for further epidemiological studies that should lead to implementation of prevention programs for toxoplasmosis.

Extruded Nucleoli of Human Dental Pulp Cells.

Background and Objectives: The dental pulp stem cells are highly proliferative and can differentiate into various cell types, including endothelial cells. We aimed to evaluate the ultrastructural characteristics of the human dental pulp cells of the permanent frontal teeth. Materials and Methods: Human adult bioptic dental pulp was collected from n = 10 healthy frontal teeth of five adult patients, prior to prosthetic treatments for aesthetic purposes. Tissues were examined under transmission electron microscopy. Results: We identified cells with a peculiar trait: giant nucleoli resembling intranuclear endoplasmic reticulum, which mimicked extrusion towards the cytoplasm. These were either partly embedded within the nuclei, the case in which their adnuclear side was coated by marginal heterochromatin and the abnuclear side was coated by a thin rim of ribosomes, or were apparently isolated from the nuclei, while still being covered by ribosomes. Conclusions: Similar electron microscopy features were previously reported in the human endometrium, as nucleolar channel system; or R-Rings induced by Nopp140. To our knowledge, this is the first report of extruded nucleolar structure in the dental pulp. Moreover, the aspect of giant extruded nucleoli was not previously reported in any human cell type, although similar evidence was gathered in other species as well as in plants.

Seroprevalence and Risk Factors of Trichinella spiralis Infection in Blood Donors from Western Romania.

Background and Objectives: Trichinellosis, a serious and sometimes fatal human disease, is a foodborne zoonotic disease with worldwide distribution caused by parasitic nematodes of the genus Trichinella. Humans are infected with Trichinella larvae through the ingestion of meat that has not been properly cooked. Romania reported most of the confirmed cases of trichinellosis among the EU countries. The aim of this cross-sectional study was to evaluate, for the first time, the seroprevalence and risk factors of Trichinella infection in blood donors from Western Romania. Materials and Methods: Serum samples of 1347 consecutive blood donors were investigated using an immunoenzymaticassay (ELISA) for the determination of specific IgG class antibodies against T.spiralis. A questionnaire interview was used to obtain information regarding the potential risk factors associated with T. spiralis infection. Mantel-Haenszel chi-squared test or the Fisher exact two-tailed test, as appropriate, were used for comparison between T. spiralis positive and T. spiralis negative blood donors. Student's t-test was used to evaluate differences between means in studied groups and body mass index was calculated by dividing weight in kilograms by height in meters squared. Statistical analysis was performed using Epi Info Version 7.2 and Stata 16.1. Results: T. spiralis IgG antibodies were detected in 2.00% (27) of 1347 consecutive blood donors. Eating raw and/or undercooked meat, from pigs or wild boars, was found to be the main risk factor (p < 0.001). Strong alcoholic drink consumption was highly associated with T. spiralis infection (p = 0.009). Trichinella seroprevalence was higher among rural residents and males. Subjects identified as Trichinella seropositive were not previously diagnosed and have not been treated for Trichinella infection with any specific therapy. Conclusions: The demonstration of T. spiralis antibodies in healthy blood donors suggests that Trichinella infection may be detected in asymptomatic individuals that were not previously diagnosed with this zoonosis.

Metformin alleviates monoamine oxidase-related vascular oxidative stress and endothelial dysfunction in rats with diet-induced obesity.

In the past decade, monoamine oxidase (MAO) with 2 isoforms, MAO-A and B, has emerged as an important source of mitochondrial reactive oxygen species (ROS) in cardio-metabolic pathologies. We have previously reported that MAO-related oxidative stress mediates endothelial dysfunction in rodent models of diabetes and diabetic patients; however, the role of MAO in the vascular impairment associated to obesity has not been investigated so far. Metformin (METF), the first-line drug in the therapy of type 2 diabetes mellitus, has been reported to elicit vasculoprotective effects via partially elucidated mechanisms. The present study was purported to assess the effects of METF on MAO expression, ROS production and vasomotor function of aortas isolated from rats with diet-induced obesity. After 24 weeks of high calorie junk food (HCJF) diet, isolated aortic rings were prepared and treated with METF (10 µM, 12 h incubation). Measurements of MAO expression (quantitative PCR and immune histochemistry), ROS production (spectrometry and immune-fluorescence) and vascular reactivity (myograph studies) were performed in rat aortic rings. MAO expression was upregulated in aortic rings isolated from obese rats together with an increase in ROS production and an impairment of vascular reactivity. METF decreased MAO expression and ROS generation, reduced vascular contractility and improved the endothelium-dependent relaxation in the diseased vascular preparations. In conclusion, METF elicited vascular protective effects via the mitigation of MAO-related oxidative stress in the rat model of diet-induced obesity.

Pleiotropic Effects of Eugenol: The Good, the Bad, and the Unknown.

Phytocompounds and medicinal herbs were used in traditional ancient medicine and are nowadays increasingly screened in both experimental and clinical settings due to their beneficial effects in several major pathologies. Similar to the drug industry, phytotherapy is interested in using nanobased delivery systems to view the identification and characterization of the cellular and molecular therapeutic targets of plant components. Eugenol, the major phenolic constituent of clove essential oil, is a particularly versatile phytochemical with a vast range of therapeutic properties, among which the anti-inflammatory, antioxidant, and anticarcinogenic effects have been systematically addressed. In the past decade, with the emerging understanding of the role of mitochondria as critical organelles in the pathophysiology of noncommunicable diseases, research regarding the role of phytochemicals as modulators of bioenergetics and metabolism is on a rise. Here, we present a brief overview of the major pharmacological properties of eugenol, with special emphasis on its applications in dental medicine, and provide preliminary data regarding its effects, alone, and included in polyurethane nanostructures, on mitochondrial bioenergetics, and glycolysis in human HaCaT keratinocytes.

Monoamine oxidase inhibition improves vascular function in mammary arteries from nondiabetic and diabetic patients with coronary heart disease.

Monoamine oxidases (MAOs) are mitochondrial enzymes with 2 isoforms that have emerged as important contributors to cardiovascular oxidative stress via the constant generation of hydrogen peroxide. The present study was purported to assess whether MAO-derived H2O2 contributes to the endothelial dysfunction in mammary arteries harvested from coronary heart disease patients with and without diabetes mellitus subjected to coronary artery bypass grafting. To this aim, the effects of MAO inhibition on vascular contractility to phenylephrine and endothelial-dependent relaxation (EDR) in response to acetylcholine were studied in vascular segments. Clorgyline (irreversible MAO-A inhibitor), selegiline (irreversible MAO-B inhibitor), and moclobemide (reversible MAO-A inhibitor) were applied in the organ bath (10 µmol/L). MAO expression was assessed by immunohistochemistry. We found a constant impairment of EDR that has been significantly attenuated in the presence of the MAO-A and MAO-B inhibitors in both groups of coronary heart disease patients. MAO-B was the dominant isoform in all human diseased vessels. In conclusion, in vitro inhibition of MAO significantly improved EDR in human mammary arteries, regardless of the presence of diabetes. These data suggest that MAO inhibitors might be useful in restoring endothelial response in clinical conditions associated with increased oxidative stress, such as coronary artery disease and diabetes.

Monoamine Oxidases as Potential Contributors to Oxidative Stress in Diabetes: Time for a Study in Patients Undergoing Heart Surgery.

Oxidative stress is a pathomechanism causally linked to the progression of chronic cardiovascular diseases and diabetes. Mitochondria have emerged as the most relevant source of reactive oxygen species, the major culprit being classically considered the respiratory chain at the inner mitochondrial membrane. In the past decade, several experimental studies unequivocally demonstrated the contribution of monoamine oxidases (MAOs) at the outer mitochondrial membrane to the maladaptative ventricular hypertrophy and endothelial dysfunction. This paper addresses the contribution of mitochondrial dysfunction to the pathogenesis of heart failure and diabetes together with the mounting evidence for an emerging role of MAO inhibition as putative cardioprotective strategy in both conditions.

Seroprevalence of Toxoplasma gondii in Western Romania.

Toxoplasma gondii is an obligate intracellular parasite that most commonly causes asymptomatic infection in immunocompetent hosts, but can have devastating consequences in congenitally infected infants and immunocompromised patients. We evaluated the seroprevalence of T. gondii in the general population in Western Romania. Sera from 304 individuals were analysed with the Pastorex Toxo test, which allows the simultaneous detection of T. gondii IgG and/or IgM antibodies. T. gondii antibodies were demonstrated in 197 individuals (64.8%) and the prevalence increased with age: 35.0% in those < 20 years versus 76.8% in those ≥ 70 years (p < 0.001). There was a higher prevalence of T. gondii antibodies in rural areas (76.9%) than in urban regions (55.3%) (p < 0.001). Our results suggest a high prevalence of T. gondii antibodies in Western Romania.

Morphological and functional aspects of sciatic nerve regeneration after crush injury.

Experimental models for the investigation of nerve regeneration are critical in studying new strategies able to promote the repair process. The aim of the present work was to characterize morphological and functional aspects of sciatic nerve regeneration after mechanical crush injury in rodents. Morphological changes were assessed after a four minutes sciatic nerve injury induced by means of a standardized compression clip. Rat nerve samples were collected before injury and after 24 hours, four days, two weeks, and four weeks after injury, respectively. In an additional group with unilateral sciatic nerve injury, animals were evaluated for four weeks using walking track analysis and the sciatic static index (SSI) measured in both rearing and normal standing position. Histological study showed important axonal degeneration at four days and axonal regeneration at four weeks after injury. We observed no significant differences between SSI in rearing and normal standing stance and a strong correlation between SSI values measured in the two positions during the evaluation period. Positive correlations were also found for the footprint parameters. Our data provide a baseline characterization of the sciatic nerve crush injury that will further allow the investigation of peripheral nerve regeneration in the presence of potential neuroprotective agents in post-traumatic nerve repair.

The association between arterial stiffness and carotid intima-media thickness in patients with known cardiovascular risk factors.

OBJECTIVES: The aim of this study was to evaluate the relationship between pulse wave velocity and carotid intima-media thickness (IMT) in patients with different cardiovascular risk factors. MATERIAL AND METHODS: This prospective study included 223 patients with at least 2 risk factors for atherosclerosis, divided based on the presence of coronary artery disease (CAD): 140 patients with angiographically documented CAD and 83 patients without CAD. The patients were compared with a control group of 74 healthy age-matched subjects (CON). We determined the following parameters: blood pressure, total cholesterol (TC), triglycerides (TG), LDL-cholesterol, HDL-cholesterol, fasting serum glucose and renal parameters: blood urea nitrogen, serum creatinine, and uric acid levels in all patients. Arterial stiffness was measured with non-invasive oscillometric Arteriograph device (Tensiomed Ltd., Budapest, Hungary) and carotid scan was performed with B-mode ultrasound. RESULTS: Patients with CAD had increased values of aortic PWV compared with patients without CAD and CON group (12.5±0.7 vs 10.9±0.6 vs 8.5±0.6 m/s, all p<0.001). The values of carotid IMT were significantly higher in CAD patients as compared to patients without CAD and CON group (1±0.05 versus 1.0±0.07 versus 0.7±0.08 mm, all p<0.001). The carotid IMT significantly correlated with aortic PWV (r=0.787, p<0.001). CONCLUSIONS: Carotid IMT and aortic PWV are useful non-invasive methods for atherosclerosis detection in patients with known cardiovascular risk factors. Both determinations in the same patient will increase the clinical relevance.

Esophageal telocytes and hybrid morphologies.

TCs (telocytes) are actually defined as stromal cells with specific long and thin prolongations, called Tp (telopodes). They have been positively identified in various tissues and we now report their presence in the esophagus. These cells were identified by TEM (transmission electron microscopy) in esophageal samples of Wistar rats (n = 5) occurring beneath the basal epithelial layer, in submucosa, closely related to smooth and striated muscular fibres, as also in the adventitia. They are closely related to mast cells, macrophages and microvessels. Hybrid morphologies of stromal cells processes were found: cytoplasmic processes continued distally in a telopodial fashion. Telopodes alone may not be sufficient, however, for a safe diagnosis of TCs in TEM. A larger set of specific standards (such as the telopodial emergence, and the size of the cell body and telopodes) should be considered to differentiate TCs from various species of fibroblasts. The morphological and ultrastructural features should distinguish between TCs and interstitial cells of Cajal in the digestive tract.

Atypical variant of lichen planus mimicking normal skin histology.

We present the case of a 69-year-old Caucasian and non-smoker patient with erythematous, itchy, violaceous lesions on the ankles, wrists, lower legs, forearms and trunk developed within 15 months. Her condition was diagnosed as prurigo and she was treated for a long period of time with antihistaminic drugs, with no resolution of lesions. In October 2008, she presented to a Private Practice of Dermatology in Timisoara. The dermatologist noticed the development of violaceous lesions on her trunk. The patient had similar lesions on the ankles, wrists, lower legs, and forearms for the last eight months. At physical examination many hyperpigmented, 1 to 6 cm, thin plaques were present on the flanks, shoulders, and infra-mammary area. There was no vaginal involvement. This eruption had a good response to topical glucocorticoids, but recurred multiple times after discontinuation of treatment. A biopsy specimen showed some histopathological features of lichen planus.

The diagnostic value of EMA expression in the renal parenchyma tumors.

Renal parenchyma tumors are a heterogeneous group of malignancies that are difficult to diagnose and classify. Immunohistochemistry begun to be routinely used for the diagnosis of these tumors. Panels of antibodies are developed for the diagnostic assessment of these tumors, which include cytokeratins, epithelial membrane antigen and vimentin. Epithelial membrane antigen (EMA) is expressed by most of the tumor cell types. Forty-seven specimens of renal parenchyma tumors were studied immunohistochemically for the expression of EMA. In the majority of the cases, clear cells carcinoma was positive for EMA (25/33, 75.70%). All of the papillary carcinomas were positive, with different staining patterns between the two subtypes. The two cases of chromophobe cells carcinomas were intensely positive with a granular cytoplasmic staining pattern. The mixed epithelial-stromal tumor was negative for EMA in both of the components. Out of the three cases of sarcomatoid carcinomas, one was negative, one was weakly positive (+1) and the last was positive (+2). Intensely positive normal tubes were caught by the tumor proliferation in the negative case and in the negative stained areas of the weakly positive case.

The diagnostic value of VEGF expression in the renal parenchyma tumors.

Tumor angiogenesis emerged as an important concept in cancer therapy over two decades ago, and was extensively studied by the discovery of VEGF family members. VEGF, also known as vascular permeability factor, is a generic name for VEGF-A, which is one of the members involved in angiogenesis. VEGF is the most important angiogenic factor, with significant effects on tumor angiogenesis. Tumor expression of VEGF was not the first angiogenesis indicator, but a growing number of studies have demonstrated that VEGF could be a prognostic factor, independent even from microvascular density, which is increased by its expression. Renal parenchyma tumors are a heterogeneous group of malignancies, difficult to classify or monitor, which prompts for the assessment of novel markers useful for the investigation of tumor histogenesis or prognostic assessment. VEGF expression in renal parenchyma tumors is poorly studied, with most of the articles published so far focusing on antiangiogenic usage in renal carcinoma therapy. The aim of this study is to detect the expression pattern of VEGF in renal parenchyma tumors by immunohistochemistry.