The role of anaemia in oxidative and genotoxic damage in transfused β-thalassaemic patients.

Redox imbalance and genotoxic damage are commonly observed in β thalassaemic patients. The aim of this study was to assess the role of anaemia in oxidative and genotoxic damage in regularly transfused thalassaemic patients, undergoing iron chelation therapy.We studied the relationships of haematological, biochemical and clinical parameters with oxidative (reactive oxygen species and 8-oxo-7,8-dihydro-2'-deoxyguanosine) and genotoxic biomarkers (Comet assay and cytokinesis-block micronucleus test) in blood samples from 105 patients. To reduce the early effect of redox-active iron, samples were collected when pharmacokinetics of the iron chelators ensured their maximum effectiveness. The transfusion regimen, cardiac and hepatic magnetic resonance imaging T2* were evaluated to characterize the patient cohort. Labile plasma iron (LPI) was also assayed.Haemoglobin level had a significant effect on ROS, %DNA in the tail and micronuclei-micronucleated cell frequency (p  0.05). Higher Hb values reduced redox imbalance. LPI, detectable in 50.5% of patients, was related to the number of apoptotic and necrotic lymphocytes (p = 0.03), demonstrating the cytotoxic effect of iron.The results highlight that an adequate transfusion regimen is essential to limit oxidative and genotoxic damage in β-thalassemic patients undergoing chelation therapy.Owing to the higher risk of cancer in the thalassaemic cohorts, specific genotoxicity/oxidative biomarkers should be monitored in order to ameliorate and formulate more personalized disease management

Peroxisome proliferator activated receptor γ is not necessary for the development of LPS-induced tolerance in macrophages

Peroxisome proliferator activated receptor-γ (PPARγ) has been reported to exert anti-inflammatory properties in endotoxic shock and sepsis. One phenomenon that alters the inflammatory response to endotoxin [lipopolysaccharide (LPS)] is endotoxin tolerance, which is caused by previous exposure to endotoxin. Here, we investigate whether changes in endogenous PPARγ function regulate this phenomenon using three different models of LPS-induced tolerance in macrophages. In a first in vitro model, previous LPS exposure of murine J774.2 macrophages suppressed tumour necrosis factor-α (TNF-α) release in response to subsequent LPS challenge. Treatment of J774.2 cells with the PPARγ inhibitor GW9662 did not alter tolerance induction because these cells were still hyporesponsive to the secondary LPS challenge. In a second ex vivo model, primary rat peritoneal macrophages from LPS-primed rats exhibited suppression of thromboxane B2 and TNF-α production, while maintaining nitrite production in response to in vitro LPS challenge. Pretreatment of rats with the PPARγ inhibitor GW9662 in vivo failed to alter the tolerant phenotype of these primary macrophages. In a third ex vivo model, primary peritoneal macrophages with conditional deletion of PPARγ were harvested from LPS-primed Cre-lox mice (Cre+/+ PPARγ−/−) and exhibited significant suppression of TNF-α production in response to in vitro LPS challenge. Furthermore, both LPS-primed PPARγ-deficient Cre+/+ PPARγ−/− mice and wild-type Cre−/− PPARγ+/+ mice exhibited reduced plasma TNF-α levels in response to a high dose of LPS in vivo. These data demonstrate that PPARγ does not play a role in the LPS-induced tolerant phenotype in macrophages

Deficiency of AMPKα1 Exacerbates Intestinal Injury and Remote Acute Lung Injury in Mesenteric Ischemia and Reperfusion in Mice

Mesenteric ischemia and reperfusion (I/R) injury can ensue from a variety of vascular diseases and represents a major cause of morbidity and mortality in intensive care units. It causes an inflammatory response associated with local gut dysfunction and remote organ injury. Adenosine monophosphate-activated protein kinase (AMPK) is a crucial regulator of metabolic homeostasis. The catalytic α1 subunit is highly expressed in the intestine and vascular system. In loss-of-function studies, we investigated the biological role of AMPKα1 in affecting the gastrointestinal barrier function. Male knock-out (KO) mice with a systemic deficiency of AMPKα1 and wild-type (WT) mice were subjected to a 30 min occlusion of the superior mesenteric artery. Four hours after reperfusion, AMPKα1 KO mice exhibited exaggerated histological gut injury and impairment of intestinal permeability associated with marked tissue lipid peroxidation and a lower apical expression of the junction proteins occludin and E-cadherin when compared to WT mice. Lung injury with neutrophil sequestration was higher in AMPKα1 KO mice than WT mice and paralleled with higher plasma levels of syndecan-1, a biomarker of endothelial injury. Thus, the data demonstrate that AMPKα1 is an important requisite for epithelial and endothelial integrity and has a protective role in remote organ injury after acute ischemic events

C-peptide, a novel inhibitor of lung inflammation following hemorrhagic shock

C-peptide is a 31-amino acid peptide cleaved from proinsulin during insulin synthesis. Initially thought to be inert, C-peptide may modulate the inflammatory response in the setting of endotoxemia and ischemia reperfusion. However, the spectrum of its biological effects is unclear. We hypothesized that exogenous administration of C-peptide would modulate pro- and anti-inflammatory signaling pathways and thereby attenuate lung inflammation in an in vivo model of hemorrhagic shock. Hemorrhagic shock was induced in male Wistar rats (aged 3–4 mo) by withdrawing blood to a mean arterial pressure of 50 mmHg. At 3 h after hemorrhage, rats were rapidly resuscitated by returning their shed blood. At the time of resuscitation and every hour thereafter, animals received C-peptide (280 nmol/kg) or vehicle parenterally. Animals were euthanized at 1 and 3 h after resuscitation. C-peptide administration at resuscitation following hemorrhagic shock ameliorated hypotension and blunted the systemic inflammatory response by reducing plasma levels of IL-1, IL-6, macrophage inflammatory protein-1α, and cytokine-induced neutrophil chemoattractant-1. This was associated with a reduction in lung neutrophil infiltration and plasma levels of receptor for advanced glycation end products. Mechanistically, C-peptide treatment was associated with reduced expression of proinflammatory transcription factors activator protein-1 and NF-κB and activation of the anti-inflammatory transcription factor peroxisome proliferator-activated receptor-γ. Our data suggest that C-peptide ameliorates the inflammatory response and lung inflammation following hemorrhagic shock. These effects may be modulated by altering the balance between pro- and anti-inflammatory signaling in the lung

C-peptide ameliorates kidney injury following hemorrhagic shock

Reperfusion injury following hemorrhagic shock is accompanied by the development of a systemic inflammatory state that may lead to organ failure. C-peptide has been shown to exert anti-inflammatory effects in sepsis and myocardial ischemia-reperfusion injury, and to ameliorate renal dysfunction in diabetic animals. Hence, we investigated the effect of C-peptide on kidney injury following hemorrhagic shock. We hypothesized that C-peptide would exert reno-protective effects by blunting inflammation. Hemorrhagic shock was induced in male rats (3–4 months old) by withdrawing blood from the femoral artery to a mean arterial pressure of 50 mmHg. Animals were kept in shock for 3h at which time they were rapidly resuscitated by returning their shed blood. At the time of resuscitation and every hour thereafter, one group of animals received C-peptide (280 nmol/kg intravenously) while another group received vehicle. Hemorrhagic shock resulted in significant rise in plasma levels of creatinine and elevated kidney neutrophil infiltration as evaluated by myeloperoxidase (MPO) activity in vehicle-treated rats in comparison with sham rats, thus suggesting kidney injury. Treatment with C-peptide significantly attenuated the rise in creatinine and kidney MPO activity when compared to vehicle group. At a molecular level these effects of C-peptide were associated with reduced expression of the c-Fos subunit and reduced activation of the pro-inflammatory kinases, extracellular signal-regulated kinase (ERK 1/2) and c-Jun N-terminal kinase (JNK) and subsequently, reduced DNA binding of activator protein-1 (AP-1) in the kidney. Thus, our data suggest that C-peptide may exert reno-protective effects following hemorrhagic shock by modulating AP-1 signaling

Affective control and life satisfaction in thalassemics

Background. Thalassemia is a chronic disease that can lead to an impact on psychological functioning and social behavior of patients. However, still little is known about the specific psychological aspects of the disease, such as the degree of tension, life satisfaction and affective control, especially in adult patients. Aim. The purpose of this study is to investigate whether patients with thalassemia have specific psychological pattern relating to the dimensions of tension, satisfaction and quality of life, management of affection. Method. We evaluated 31 patients with thalassemia major and intermedia (19 women and 12 men) aged between 18 and 50 years (M = 34 + 16), belonging to the Complex Unit of Medical Genetics. For the evaluation were used the Profile of Mood States (POMS), the Quality of Life Enjoyment and Satisfaction Questionnaire (Q-LES-Q) and the Rorschach test. Results. The findings show an inverse relationship between the levels of self-reported tension and the affective control indicators at Rorschach. Life satisfaction, instead, seems to vary according to the severity of the disease - major vs. intermediate - and the type of therapy. Conclusions. An understanding of the psychological mechanisms involved in thalassemia, both self-reported and projective, can contribute to a wider patient takeover, by considering the subjective aspects related to the psychological and socioemotional well-being, fundamental in the care compliance

Affective control and life satisfaction in thalassemics

Background. Thalassemia is a chronic disease that can lead to an impact on psychological functioning and social behavior of patients. However, still little is known about the specific psychological aspects of the disease, such as the degree of tension, life satisfaction and affective control, especially in adult patients.Aim. The purpose of this study is to investigate whether patients with thalassemia have specific psychological pattern relating to the dimensions of tension, satisfaction and quality of life, management of affection.Method. We evaluated 31 patients with thalassemia major and intermedia (19 women and 12 men) aged between 18 and 50 years (M = 34 + 16), belonging to the Complex Unit of Medical Genetics. For the evaluation were used the Profile of Mood States (POMS), the Quality of Life Enjoyment and Satisfaction Questionnaire (Q-LES-Q) and the Rorschach test.Results. The findings show an inverse relationship between the levels of self-reported tension and the affective control indicators at Rorschach. Life satisfaction, instead, seems to vary according to the severity of the disease - major vs. intermediate - and the type of therapy.Conclusions. An understanding of the psychological mechanisms involved in thalassemia, both self-reported and projective, can contribute to a wider patient take-over, by considering the subjective aspects related to the psychological and socio-emotional well-being, fundamental in the care compliance.Antecedentes: La Talasemia es una enfermedad crónica que puede conducir a un impacto en el funcionamiento psicológico y el comportamiento social de los pacientes. Sin embargo, todavía se sabe poco sobre los aspectos psicológicos específicos de esta enfermedad, tales como el grado de tensión, satisfacción con la vida y el control afectivo, sobre todo en pacientes adultos.Objetivo: El propósito de este estudio, es investigar si los pacientes con Talasemia tienen un patrón psicológico específico relacionado con las dimensiones de tensión, la satisfacción, calidad de vida y el manejo del afecto.Método: 31 pacientes con Talasemia mayor e intermedia fueron evaluados: (19 mujeres y 12 hombres) con edades entre 18 y 50 años (M= 34+16), pertenecientes a la Unidad de Complejo de Genética Médica. Para la evaluación fueron utilizados el Perfil de Estados de Ánimo (POMS), la calidad de vida: Satisfacción y placer (Q-LES-Q) y el test de Rorschach.Resultados: Los resultados muestran una relación inversa entre los niveles de tensión de auto-reporte y los indicadores de control afectivo en el test de Rorschach. La satisfacción con la vida, por el contrario parece variar según la gravedad de la enfermedad: major vs. intermedio - y el tipo de terapia.Conclusiones: la comprensión de los mecanismos psicológicos implicados en la talasemia, tanto en el auto-reporte como en lo proyectivo, puede contribuir a una toma de control más amplia del paciente, considerando los aspectos subjetivos relacionados con el bienestar psicológico y socio-emocional, fundamentales en el cumplimiento del cuidado