Coralia Cotoraci

Active Caspases-8 and -3 in Circulating Human Erythrocytes Purified on Immobilized Annexin-V: A Cytometric Demonstration

Human red blood cells (RBCs) have a normal life span of 120 days in vivo and might be primed in vitro to die in response to apoptotic stimuli through a caspase‐independent pathway. It is well known that, in vivo, aging RBCs externalize phosphatidylserine residues but is unknown whether these cells express active caspases at this stage. We isolated RBCs expressing phosphatidylserine on their surface from human blood by applying an original method of affinity chromatography using annexin‐V fixed on gelatin or on magnetic beads. The isolated RBCs were then analyzed by flow cytometry for morphological changes (dot‐plot forward scatter versus side scatter), phosphatidylserine externalization (annexin‐V test), cell viability (calcein‐AM test), and caspase activities using fluorescent substrates specific for caspases‐3 and −8. In addition, cells were systematically visualized using phase contrast, fluorescence, and confocal microscopy. We found that the population of RBCs fixed on annexin‐V is a mixture of discocytes and shrunken cells. This annexin‐V‐positive population showed a dramatic loss of viability based on esterase activity determination (calcein‐AM test). Moreover, we demonstrated that circulating RBCs express both active caspases‐8 and −3 in half of the annexin‐V‐positive cells. All of these results were confirmed by phase contrast, fluorescence, and confocal microscopy. Our results demonstrate active caspases in RBC isolated from blood suggesting that caspases may participate in the regulation of in vivo RBC half‐life. This finding open the door to fruitful investigations in the field of RBC pathology.

A cytometric study of the red blood cells in Gaucher disease reveals their abnormal shape that may be involved in increased erythrophagocytosis

Background: Gaucher disease is a sphingolipidosis caused by a deficiency of the enzyme glucocerebrosidase. Macrophages transform into pathogenic Gaucher cells following the phagocytosis of red blood cells (RBCs) and subsequent accumulation of glucosylceramide. Enhanced erythrophagocytosis is one feature of the disease indicating abnormal macrophage‐RBC interactions. We hypothesized that the erythrophagocytosis observed in Gaucher disease may be at least partly due to abnormalities in the RBCs themselves.

Protective effects of silymarin on epirubicin-induced mucosal barrier injury of the gastrointestinal tract

Abstract Mucositis is a serious disorder of the gastrointestinal tract that results from cancer chemotherapy. We investigated the protective effects of silymarin on epirubicin-induced mucosal barrier injury in CD-1 mice. Immunohistochemical activity of both pro-apoptotic Bax and anti-apoptotic Bcl-2 markers, together with p53, cyt-P450 expression and DNA damage analysis on stomach, small intestine and colon were evaluated. Our results indicated stronger expression for cyt P450 in all analyzed gastrointestinal tissues of Epi group, which demonstrate intense drug detoxification. Bax immunopositivity was intense in the absorptive enterocytes and lamina connective cells of the small intestine, surface epithelial cells of the stomach and also in the colonic epithelium and lamina concomitant with a decreased Bcl-2 expression in all analyzed tissues. Epirubicin-induced gastrointestinal damage was verified by a goblet cell count and morphology analysis on histopathological sections stained for mucins. In all analyzed tissues, Bax immunopositivity has been withdrawn by highest dose of silymarin concomitant with reversal of Bcl-2 intensity at a level comparable with control. p53 expression was found in all analyzed tissues and decreased by high dose of silymarin. Also, DNA internucleosomal fragmentation was observed in the Epi groups for all analyzed tissues was almost suppressed at 100 mg/kg Sy co-treatment. Histological aspect and goblet cell count were restored at a highest dose of Sy for both small and large intestine. In conclusion, our findings suggest that silymarin may prevent cellular damage of epirubicin-induced toxicity and was effective in reducing the severity indicators of gastrointestinal mucositis in mice.

Protective effects of silymarin against bisphenol A-induced hepatotoxicity in mouse liver

Bisphenol A (BPA) is an endocrine-disrupting chemical released into the environment, with severe consequences for human health, including metabolic syndrome and associated pathological conditions. Due to limited information on BPA-induced hepatotoxicity, the present study focused on investigating the association between BPA-induced toxicity and inflammatory markers in the liver, and how these injuries may be alleviated using the natural agent silymarin, a flavonoid with antioxidant properties obtained from Silybum marianum. Administration of BPA to male CD-1 mice for 10 days caused a significant increase in the number of cells immunopositive for interleukin 6 and tumor necrosis factor-α, pro-inflammatory cytokines that mediate the hepatic inflammatory response. Treatment with 200 mg/kg of silymarin concurrently with BPA for 10 days resulted in a diminished level of pro-inflammatory cytokines and in significantly reduced ultrastructural injuries. Additionally, silymarin was able to restore the significantly decreased glycogen deposits observed following BPA exposure to normal levels, thus favoring hepatic glycogenesis. This study represents the first report of silymarin ability to reduce hepatic lesions and to counteract inflammation caused by BPA in mice. A dose of 200 mg/kg silymarin was sufficient to induce a protective effect against structural and ultrastructural injuries induced by BPA and to lower the levels of pro-inflammatory cytokines observed in murine liver tissue following exposure to BPA.

Evaluation of Hepatotoxicity with Treatment Doses of Flucytosine and Amphotericin B for Invasive Fungal Infections

Invasive fungal infection is a well-known cause of morbidity and mortality in immunocompromised patients. In this study we aimed to evaluate the hepatotoxicity induced by combined therapy of flucytosine and amphotericin B, at three different doses administered to mice for 14 days: 50 mg/kg flucytosine and 300 μg/kg amphotericin B; 100 mg/kg flucytosine and 600 μg/kg amphotericin B; 150 mg/kg flucytosine and 900 μg/kg amphotericin B. Liver injuries were evaluated by analysis of optic and electron microscopy samples, changes in TNF-α, IL-6, and NF-κB inflammation markers levels of expression, and evaluation of mRNA profiles. Histological and ultrastructural analysis revealed an increase in parenchymal and portal inflammation in mice and Kupffer cells activation. Combined antifungal treatment stimulated activation of an inflammatory pathway, demonstrated by a significant dose-dependent increase of TNF-α and IL-6 immunoreactivity, together with mRNA upregulation. Also, NF-κB was activated, as suggested by the high levels found in hepatic tissue and upregulation of target genes. Our results suggest that antifungal combined therapy exerts a synergistic inflammatory activation in a dose-dependent manner, through NF-κB pathway, which promotes an inflammatory cascade during inflammation. The use of combined antifungal therapy needs to be dose limiting due to the associated risk of liver injury, especially for those patients with hepatic dysfunction.

Haematological Patients’ Perception of Their Quality of Life

The therapeutic management of haematological patients dramatically changes the lives of patients. Quality of life is a modern concept which tries to help the patient and refocuses doctors on improving not only the patient’s symptoms and disease itself, but also the patient’s overall well-being. This chapter will highlight the most important factors which can modify the onco-haematological patient’s quality of life, stressing the importance of biological, psychological, and social factors. In identifying these factors, we used two standard questionnaires which evaluate the quality of life on various fields. The results showed that physical status, mental health, and social factors influence patient quality of life.

Role of Optimal Medication Given to Patients with Hypertension and Ischemic Heart Disease Prior to an Acute Coronary Syndrome

Introduction Administering optimal cardiovascular medication (OCM) to patients with hypertension (HBP) and ischemic heart disease (IHD) lowers cardiovascular morbidity and mortality. The main objective of this study was to compare in-hospital cardiac mortality among patients with HBP and/or IHD, treated or untreated with OCM, who developed a first episode of acute coronary syndrome (ACS). Methods The study was carried out retrospectively and included patients admitted with a first episode of ACS between 2013 and 2016. The patients were divided into three groups: those with HBP, IHD, and a history of HBP + IHD. Patients were then divided into two subgroups: subgroup A consisted of patients undergoing optimal anti-ischemic and/or antihypertensive therapy, while subgroup B consisted of patients without OCM. Results This analysis comprised 1096 patients. Mean age was 64.3 ± 18 years. There were 581 patients in subgroup A – 53%, and 515 patients in subgroup B – 47%. Total cardiac mortality was 9.98%, different depending on the groups and subgroups studied: HBP group total – 7%, subgroup A – 5.1%, significantly lower compared to subgroup B – 9.4% (p = 0.05); IHD group total – 12.2%, subgroup A – 9.07%, significantly lower compared to subgroup B – 15.8% (p = 0.05); HBP + IHD group total – 14.35%, subgroup A – 9.9%, significantly lower compared to subgroup B – 18.8% (p = 0.05). Conclusions The lack of OCM in patients with HBP and/or IHD is correlated to a significant increase in in-hospital cardiac mortality among patients who develop a first-episode ACS.