Deposition In Heart Research Articles

Correlation between plasma biochemical parameters and cardio-hepatic iron deposition in thalassemia major patients

Introduction Major Thalassemia patients suffer from iron overload and organ damage, especially heart and liver damage. Early diagnosis and treatment with a chelator can reduce the complications and mortality of iron overload. Therefore, we aimed to investigate the biochemical and hematological predictors as an alternative and indirect indicator of iron deposition in heart and liver cells in comparison with the MRI T2* method as the gold standard. Material and method MRI T2* was evaluated in the heart and liver tissues of 62 major beta-thalassemia patients undergoing regular transfusion and chelator therapy. Biochemical and hematological factors were also measured, including serum ferritin, serum electrolytes, liver enzymes, hemoglobin, blood glucose, and serum magnesium. The correlation between these factors was assessed using statistical evaluations. Result Serum ferritin had a positive and significant correlation with liver siderosis based on MRI T2* (p-value = .015), and no significant association was observed with cardiac siderosis (p-value = .79). However, there was a significant positive correlation between cardiac iron deposition and fasting blood sugar level (p-value = −.049), and plasma level of liver enzymes (alanine aminotransferase (ALT) (p-value = .001), aspartate aminotransferase (AST ((p-value = .01)). Moreover, there was a significant negative correlation between cardiac iron overload and plasma magnesium level (p-value = .014). According to MRI T2*, there was no significant correlation between cardiac and hepatic iron overload (p value = .36). Conclusion An increase in blood sugar or liver enzymes and a decrease in serum magnesium was associated with an increase in cardiac iron overload based on MRI T2*. Liver iron overload based on MRI T2* had a significant correlation with serum ferritin.

Preclinical evaluation of Tc-99m p5+14 peptide for SPECT detection of cardiac amyloidosis.

Amyloid deposition is a cause of restrictive cardiomyopathy. Patients who present with cardiac disease can be evaluated for transthyretin (TTR)-associated cardiac amyloidosis using nuclear imaging with 99mTc-labeled pyrophosphate (PYP); however, light chain-associated (AL) cardiac amyloid is generally not detected using this tracer. As an alternative, the amyloid-binding peptide p5+14 radiolabeled with iodine-124 has been shown to be an effective pan-amyloid radiotracer for PET/CT imaging. Here, a 99mTc-labeled form of p5+14 peptide has been prepared to facilitate SPECT/CT imaging of cardiac amyloidosis. A synthesis method suitable for clinical applications has been used to prepare 99mTc-labeled p5+14 and tested for peptide purity, product bioactivity, radiochemical purity and stability. The product was compared with99mTc-PYP for cardiac SPECT/CT imaging in a mouse model of AA amyloidosis and for reactivity with human tissue sections from AL and TTR patients. The 99mTc p5+14 tracer was produced with >95% yields in radiopurity and bioactivity with no purification steps required and retained over 95% peptide purity and >90% bioactivity for >3 h. In mice, the tracer detected hepatosplenic AA amyloid as well as heart deposits with uptake ~5 fold higher than 99mTc-PYP. 99mTc p5+14 effectively bound human amyloid deposits in the liver, kidney and both AL- and ATTR cardiac amyloid in tissue sections in which 99mTc-PYP binding was not detectable. 99mTc-p5+14 was prepared in minutes in >20 mCi doses with good performance in preclinical studies making it suitable for clinical SPECT/CT imaging of cardiac amyloidosis.

Correlations between pathological deposition and non-invasive diagnostic modalities like 99mTc-PYP scintigraphy, cardiac magnetic resonance, GLS in patients with transthyretin cardiac amyloidosis

Abstract Background Although wild-type transthyretin amyloid cardiomyopathy (ATTRwt-CM) was previously considered a rare disease, recent diagnostic imaging modalities have revealed that it is considerably underdiagnosed among elderly patients with heart failure. The severity of CM is thought to be related to the extent of amyloid deposition in heart. 99mTc-labeled pyrophosphate (99mTc-PYP) scintigraphy, cardiovascular magnetic resonance (CMR), global longitudinal strain (GLS) provide diagnostic and prognostic information in ATTRwt-CM. However, the relevance of these imaging modalities and their association with cardiac amyloid load has not been fully evaluated. Purpose The aim of study was to elucidate the associations between pathological amyloid load and cardiac retention evaluated by 99mTc-PYP scintigraphy, CMR, GLS in patients with ATTRwt-CM. Method Cardiac amyloid load was calculated as (amyloid deposition area/ total myocardium area)×100 using endomyocardial biopsy specimen. Cardiac retention was quantified by heart to contralateral (H/CL) ratio by 99mTc-PYP scintigraphy. Native T1 and extracellular volume (ECV) were obtained by CMR. GLS was analyzed using the 2D echo at the time of diagnosis. Result The mean cardiac amyloid load was 23.0±15.2% (n=57) and correlation with H/CL ratio (1.94±0.36 n=57), native T1 (1426.7±52.5 n=57), ECV (57.9±12.9 n=54), GLS (−9.1±2.4 n=57) were positive (r=0.375 p=0.004, r=0.496 r=0.304 p<0.001, r=0.304 p=0.025, r=0.473 p<0.001). Conclusion Increased cardiac amyloid load correlated with an increased 99mTc-PYP positivity, native T1, ECV, and an impaired GLS. These results suggest that imaging parameters may reflect histological and functional changes due to amyloid deposition in the myocardium. Funding Acknowledgement Type of funding sources: None.

Selenium, a dietary-antioxidant with cardioprotective effects, prevents the impairments in heart rate and systolic blood pressure in adolescent rats exposed to binge drinking treatment

ABSTRACT Background Binge drinking (BD) during adolescence is related to cardiovascular alterations. Selenium (Se) is an essential trace element with antioxidant, anti-inflammatory and antiapoptotic properties, essential for correct heart function. Objectives To study the protective cardiovascular effects of selenium in adolescent rats exposed to a BD-like procedure. Methods 32 adolescent male rats exposed to an intraperitoneally BD-like model or not, and supplemented with 0.4ppm of selenite or not, were divided into 4 groups: control, alcohol, control-selenium and alcohol-selenium. Blood pressure and heart rate (HR) were determined after experimentation. Se deposits, oxidative balance and the expression of glutathione peroxidases (GPxs), NF-kB and caspase-3 were measured in the heart. Also, DNA instability in rat lymphocytes and serum vascular markers were determined. Statistical analysis was performed with the ANOVA model. Results The BD-like model depleted Se heart deposits (p < .01), decreased GPx activity (p < .01) and GPx1 (p < .001) and GPx4 (p < .05) expression, increased NF-kB (p < .01), caspase-3 (p < .001) expression, and generated oxidation in myocytes. Outside the heart, the BD-like model caused double-strand breaks in lymphocyte DNA and increased all the vascular markers measured. These cardiovascular alterations were related to higher systolic (p < .001) and diastolic (p < .05) blood pressure and HR (p < .05). In the heart, Se supplementation in BD-exposed rats significantly increased Se deposits (p < .001) and improved oxidative balance and vascular damage, including increased GPxs and decreased NF-kB and caspase-3 activation, consequently decreasing systolic (p < .05) blood pressure and HR (p < .01). Conclusions Se supplementation presents cardioprotective effects since it reversed HR and systolic blood pressure observed in BD-exposed adolescent rats.

Iron, Insulin and Coronary Heart Disease Emphasis on the Generation of Sudden Death and Occult Diabetes (And Why Donating Blood Can Reduce Cardiovascular Death)

Dysregulated glucose metabolism promote inflammation in monocytes and macrophages from patients with atherosclerotic coronary artery disease. Men with metabolic syndrome are at increased risk for sudden cardiac death, and the incident sudden death is not explained by obesity or traditional cardiovascular risk factors (Kurl, 2016), but the ingested nutritional iron. Individuals with increased abdominal adiposity exhibit an increased risk of heart failure, in spite of there are oweweigtht or not (Cavalera, 2014), because the insulin resistance contribute to increased myocardial fibrosis in the absence of hypertension (Quilliot, 2005). Iron overload, and plasma viscosity contributes to cardiovascular risk in the general population, particularly in men (Van der A, 2005; Junker, 1998). Iron influences glucose metabolism, even in the absence of significant iron overload, and its reduction may alleviate coronary heart disease and reduced or prevent their complications: High stored or free iron levels (measured by serum ferritin or catalytic iron concentrations) elevate risk for development of coronary atherosclerosis, because labile iron accelerates endothelial dysfunction and originates oxidative injury that promotes systemic and vascular inflammation, phrothrombotic conditions and insulin resistance (Williams, 2002). High serum ferritin is strongly and independently associated with acute myocardial infarction and constitutes a novel risk factor in acute sudden event (Holay, 2012). Iron plays a direct and causal role in diabetes pathogenesis mediated both by β-cell failure and insulin resistance (Simcox, 2013). In the general population, body iron stores are positively associated with the development of glucose intolerance, type 2 diabetes and gestational diabetes. In this way, blood donation significant drops in the incidence of cardiovascular events, as well as in procedures such as percutaneous transluminal coronary angioplasty and coronary artery bypass grafting (Holsworth, 2013): frequent blood donations decreased iron stores in healthy volunteers, improving insulin sensitivity and hemodynamic parameters. Iron and oxygen-derived free radicals are important in the pathogenesis of postischemic reperfusion injury and contributes substantially to endothelial dysfunction in acute coronary syndromes (Chekanov, 2002; Duffy, 2001) , and a high iron diet potentially increase ischemic damage induced by transient ischemia and early reperfusion (García-Yébenes, 2012) in animals and humans. Iron, hyperinsulinemia, and hyperglycemia act in concert to up regulate free-radical reactions (Facchini, 2000) and this metal excess accelerated the development of atherosclerosis and its accumulation may promotes illness, particularly, ischemic cardiovascular diseases. Insulin resistance in macrophages promotes formation of a necrotic core in atherosclerotic plaques by enhancing macrophage apoptosis, and exposure it to circulating blood in the event of plaque rupture can precipitate thrombosis, leading to unstable angina pectoris, myocardial infarction and sudden death (Rask.Madsen, 2012, rev). In humans phlebotomy slows progression of peripheral vascular disease and blood donation lowers significantly the risk of myocardial infarction (Salonen, 1998), particularly in insulin-resistant subjects. On the contrary, exogenous iron into healthy individuals provoked endothelial dysfunction accompanied by increased generation of superoxide radical in whole blood (Vinchi, 2014, rev). A causal relationship between pre-diabetes and cardiovascular disease exist (Ford, 2010, rev). Humans lack effective mechanisms to excrete excess iron, and excessive dietary iron uptake cause iron deposition in heart, and pancreas (Kulaksis, 2008) leading to sudden death and occult diabetes mellitus. Iron play an underappreciated role in the development of insulin resistance and insulin resistance-induced heart failure. In a chronic and acute way, Insulin resistance is an early and major factor in the development of heart failure and acute iron induced insulin resistance in cardiomyocytes (Sung, 2019). 1- Introduction 2- Catalytic Iron; Irreversible Oxidizer 3- Excess Hemoglobin and Cardiocerebrovascular Pathology 4- Free Iron as a promoter of Ischemic Heart Disease 5- Excess Ferritin and pathogenesis of Endothelial Dysfunction 6- Hyperinsulinemia as a Sudden Death Promoter. The determining role of Iron 7- Iron accumulated in excess and pathogenesis of Diabetes: High hemoglobin versus High Ferritin 8- Iron and atherosclerosis. The evidence 9- Blood Donation. Cardiovascular Pathology Protector

Combinatorial inhibition of Angiotensin converting enzyme, Neutral endopeptidase and Aminopeptidase N by N-methylated peptides alleviates blood pressure and fibrosis in rat model of dexamethasone-induced hypertension

Angiotensin converting enzyme (ACE), neutral endopeptidase (NEP) and aminopeptidase N (APN) are responsible for generation of vasoactive peptides that regulates vasoconstriction, vasodilation and natriuresis, which altogether regulate blood pressure. Cumulative inhibition of ACE, NEP and APN effectively blocks the progression of respective pathways. In this study, N-methylated peptide inhibitors F-N(Me)H-L, V-N(Me)F-R and R-N(Me)V-Y were synthesized against ACE, NEP and APN respectively, using their respective physiological substrates. F-N(Me)H-L inhibited ACE activity with an IC50 of 83 nmol/L, V-N(Me)F-R inhibited NEP activity with an IC50 of 1.173 μmol/L and R-N(Me)V-Y inhibited APN activity with an IC50 of 3.94 nmol/L respectively. Further, the anti-hypertensive effect of N-methylated peptides was evaluated using rat model of dexamethasone-induced hypertension. Individual peptides and their cocktail treatment were started from day 6 of the study period and blood pressure was measured on every alternate day during 15 day study. Administration of F-N(Me)H-L (138 ± 3 mmHg) and cocktail of all the three peptides at a dose of 100 mg/kg significantly reduced systolic blood pressure (SBP) compared to dexamethasone group (SBP of Groups-dexamethasone; (167 ± 5 mmHg), F-N(Me)H-L (138 ± 3 mmHg), and Cocktail (122 ± 3 mmHg). Anti-hypertensive, anti-hypertrophic and anti-fibrotic effects of N-methylated peptides and cocktail was further reflected by the decreased levels of circulating Ang II and increased ANP levels in sera of hypertensive rats along with decrease in collagen deposition in heart and kidney. Though, ACE inhibition is adequate to reduce SBP, targeting NEP and APN along with ACE is beneficial in tackling hypertension and associated fibrosis of heart.

P897Diagnostic utility of tissue biopsy in transthyretin cardiac amyloidosis diagnosed by non-invasive diagnostic criteria

Abstract Background In the recent past, transthyretin cardiac amyloidosis (TTR-CA) is recognized as an important cause of heart failure with preserved ejection fraction, and diagnostic utility of 99mTc-pyrophosphate (99mTc-PYP) scintigraphy in TTR-CA has been established. However, pathological evaluation is still the basis to confirm diagnosis. Subcutaneous tissue and gastrointestinal tract biopsy are useful examinations alternative to endomyocardial biopsy. Nevertheless, the positive ratio of amyloid deposition in these organs is not fully evaluated based on recent non-invasive diagnostic criteria. Purpose Our aim was to evaluate the diagnostic sensitivity of tissue biopsy in TTR-CA patients diagnosed by 99mTc-PYP scintigraphy. Methods We retrospectively evaluated 124 consecutive TTR-CA patients [wild-type (ATTRwt): 90, hereditary (hATTR): 34] who were diagnosed by positive findings of 99mTc-PYP scintigraphy (visual score 2 or 3) between June 2002 and January 2019 at our institute. Results A total of 114 patients underwent tissue biopsy at least one organ. Amyloid was detected on Congo red staining of subcutaneous tissue (43/97: 44%), gastrointestinal tract (52/78: 67%) and heart (78/79: 99%), respectively. Among 70 patients who underwent both subcutaneous tissue and gastrointestinal tract biopsy, amyloid was detected at least one specimen in 57/70 (81%). Compared to hATTR CA patients, ATTRwt CA patients had lower positive ratio of subcutaneous tissue (81% vs. 27%) and gastrointestinal tract (73% vs. 63%) biopsy. Conclusion Non-invasive diagnostic criteria of TTR-CA had excellent sensitivity of amyloid deposition in heart. The diagnostic sensitivity of subcutaneous tissue and gastrointestinal tract biopsy was lower in ATTRwt CA compared to hATTR CA. Combination of subcutaneous tissue and gastrointestinal tract biopsy was useful for pathological confirmation of amyloid deposition without endomyocardial biopsy.

Role of Tissue Iron in Development of Cardiovascular Delayed Effects of Acute Radiation Exposure

The threat of radiation exposure from terrorist activity, warfare, or accidents creates an urgent need to develop agents to mitigate the acute and chronic effects of high dose rate total body irradiation (TBI). High level short‐term radiation exposure results in rapid organ injury leading to development of the acute radiation syndrome (ARS), which if untreated will result in death within weeks. Survivors of ARS will experience the delayed effects of acute radiation exposure (DEARE), which include development of chronic pathology and dysfunction of multiple organ systems months to years following exposure. However, the factors and mechanisms that regulate the onset and progression of cardiovascular DEARE are unclear. The goal of the current study was to determine the possible role for abnormal iron metabolism in the development of cardiac DEARE, especially vascular pathology. Studies utilized the ARS mouse model with gamma radiation exposure at 853 cGy (LD50) and time points from 1 day to 4 months post‐TBI.Serum iron was elevated at 1 day post‐TBI, peaked at 2 weeks, and returned to non‐irradiated control values by 4 weeks post‐TBI. A similar trend was seen for transferrin saturation, and both results correlated inversely with red blood cell number, hematocrit, and hemoglobin concentrations. Perls' Prussian Blue staining was used to detect iron (hemosiderin) deposition in heart paraffin‐embedded tissue sections, and myocardial iron was present as early as 2 weeks post‐TBI. Pretreatment of mice with the iron chelator deferiprone in drinking water decreased myocardial iron deposition (P=0.055), but not serum iron, at 2 weeks. A histological assessment of coronary artery endothelial cell (EC) density showed intimal nuclei per unit area significantly decreased as early as two weeks post‐TBI vs. non‐irradiated controls (P&lt;0.05), and the decrease persisted to 4 months post‐TBI. EC density in deferiprone‐treated mice was significantly increased at 2 and 4 weeks post‐TBI compared to irradiated non‐treated mice (P&lt;0.03). Administration of the ferroptosis inhibitor ferrostatin‐1 via daily i.p. injection for 2 weeks post‐TBI resulted in no change in coronary artery EC density compared to vehicle controls. Taken together, the results suggest the presence of tissue iron contributes to endothelial cell loss occurring early following TBI and is a significant event impacting the development of DEARE. This information may facilitate development of novel medical countermeasures to prevent or mitigate cardiovascular DEARE‐related pathophysiology.Support or Funding InformationNIH 1U01AI107340‐01 (C. Orschell), IUPUI Research Support Funds Grant (S. Miller)This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

The edible native Australian fruit, Davidson’s plum (Davidsonia pruriens), reduces symptoms in rats with diet-induced metabolic syndrome

Abstract Davidson’s plum is a native Australian fruit used traditionally as food and medicine. The fruit contains anthocyanins as glucosides and sambubiosides of cyanidin and peonidin (691 mg/100 g of dried pulp) with rutin and quercetin glycosides (193 mg/100 g). Dietary supplementation of Davidson’s plum at approximately 8 mg anthocyanins/kg/day in rats attenuated the signs of metabolic syndrome induced by high-carbohydrate, high-fat diet. Davidson’s plum reduced visceral fat accumulation, total abdominal fat weight, size of retroperitoneal adipocytes, and plasma triglycerides and non-esterified fatty acids, normalised blood pressure, reduced left ventricular stiffness, decreased infiltration of inflammatory cells in both left ventricle and liver, decreased collagen deposition in heart, and reduced both fat vacuoles in liver and obesity-induced degeneration of knee cartilage. There were no changes in glucose tolerance with treatment. Davidson’s plum reduced colonic Clostridiaceae spp, and increased Turicibacter spp. and Akkermansia muciniphila. Our findings indicate that Davidson’s plum is a potential complementary treatment for metabolic syndrome.

Cardioprotective effects of nanoceria in a murine model of cardiac remodeling

Isoproterenol (ISO), a synthetic β1 adrenergic agonist is a well-known agent to be associated with severe cardiotoxicity manifested as marked myocardial necrosis and fibrosis. Oxidative stress plays a crucial role in mediating ISO induced cardiotoxicity. In present study, we have investigated the possible protective effect of nanoceria (NC) in ISO induced cardiac injury. We have given long duration exposure (a total of 10 days) of low dose ISO (20 mg/kg/day) to investigate the protective effects of NC in chronic cardiac injury model. ISO (20 mg/kg/day for 10 days) produced cardiac injury as evident by increased plasma LDH and CK-MB, AST, ALT, cardiac hypertrophy, severe myocardial fibrosis (MF) and significantly higher levels of cytokines, IL-6, TGF-β and TNF-α. Interestingly, the treatment with NC (0.2 and 2 mg/kg) abrogated cardiotoxicity symptoms and provided protection from ISO induced cardiac injury. The results from present study demonstrated strong evidences of cardioprotective effects of NC as shown by reduction in the levels of LDH (p < 0.05 at 2 mg/kg) and CK-MB (p < 0.05 at 2 mg/kg). In addition, NC reduced oxidative stress parameters MDA (p < 0.05 at 2 mg/kg) and enhanced GSH levels which is physiological antioxidant (p < 0.01 at both doses). Further, NC exhibited promising anti-inflammatory activity and curbed the levels of cytokines (p < 0.05 at 0.2 mg/kg and p < 0.001 for IL-1β and p < 0.001 at both doses for IL-6). In addition, NC also reduced the levels of pro-fibrotic cytokine, TGF-β (p < 0.05 at 2 mg/kg) and helped in reduction of collagen deposition in heart thereby, preventing the myocardial remodeling. Our results strongly suggested that NC might be of potential use as a cardioprotective agent.

Cardioprotective effect of Amaranthus tricolor extract in isoprenaline induced myocardial damage in ovariectomized rats

Red spinach (Amaranthus tricolor) has been reported to possess many benefits and medicinal properties and used as a part of traditional medicine in Ayurveda and Siddha. The aim of the study was to investigate the effects of Amaranthus tricolor on isoproterenol-induced oxidative stress, fibrosis, and myocardial damage in ovariectomized rats. Ovariectomy surgery was conducted to remove both ovaries from the rats. After recovery, rats were administered with ISO subcutaneously (50 mg/kg) twice a week and were treated with ethanolic extracts of A. tricolor. This investigation showed that the level of oxidative stress markers was significantly increased while the superoxide dismutase (SOD) activity decreased in ISO administered ovariectomized rats. A. tricolor extract and atenolol treatment prevented the rise of malondialdehyde, nitric oxide and advanced protein oxidation product. Moreover, elevated activities of AST, ALT, and CK-MB enzymes were also lowered by both atenolol and A. tricolor treatment. Increased uric acid and creatinine levels were also normalized by atenolol, and A. Tricolor treatment in ISO administered ovariectomized rats. ISO-induced ovariectomized rats also showed massive inflammatory cell infiltration, fibrosis and iron deposition in heart compared to sham rats. Atenolol and A. tricolor treatment prevented the inflammatory cells infiltration, fibrosis, and iron deposition. These results suggest that A. tricolor treatment may protect against ISO administered myocardial infarction in ovariectomized rats probably by preventing inflammation, oxidative stress, and fibrosis. Further research is warranted to examine molecular mechanism of cardioprotective effect of A. tricolor.

Identification and characterization of Xenopus tropicalis common progenitors of Sertoli and peritubular myoid cell lineages.

ABSTRACTThe origin of somatic cell lineages during testicular development is controversial in mammals. Employing basal amphibian tetrapod Xenopus tropicalis we established a cell culture derived from testes of juvenile male. Expression analysis showed transcription of some pluripotency genes and Sertoli cell, peritubular myoid cell and mesenchymal cell markers. Transcription of germline-specific genes was downregulated. Immunocytochemistry revealed that a majority of cells express vimentin and co-express Sox9 and smooth muscle α-actin (Sma), indicating the existence of a common progenitor of Sertoli and peritubular myoid cell lineages. Microinjection of transgenic, red fluorescent protein (RFP)-positive somatic testicular cells into the peritoneal cavity of X. tropicalis tadpoles resulted in cell deposits in heart, pronephros and intestine, and later in a strong proliferation and formation of cell-to-cell net growing through the tadpole body. Immunohistochemistry analysis of transplanted tadpoles showed a strong expression of vimentin in RFP-positive cells. No co-localization of Sox9 and Sma signals was observed during the first three weeks indicating their dedifferentiation to migratory-active mesenchymal cells recently described in human testicular biopsies.

Continued improvement in overall survival in elderly multiple myeloma patients after 2008; a population based study from the Danish Multiple Myeloma Registry

Graduate School of Health Sciences, Kumamoto University; Department of Morphological and Physiological Sciences, Kumamoto University; Faculty of Life Sciences, Department of Neurology, Kumamoto University; Diagnostic Unit for Amyloidosis, Kumamoto University Hospital; Faculty of Life Sciences, Department of Hematology, Kumamoto University; Department of Surgical Pathology, Kumamoto General Hospital; Division of Informative Clinical Sciences, Kumamoto University Introduction: AL amyloidosis is a disease featuring deposition of Ig light chain (LC). Diagnosis is determined if the presence of LC in amyloid deposits is detected with anti-LC chain antibodies. Poor prognosis cases, like detected amyloid deposits in heart, require the prompt diagnoses. However, in several cases, the existence of LC is not detected within amyloid deposits with immunohistochemistry (IHC) using conventional anti-LC antibodies although monoclonal lambda light chain in serum or urine was proved. In such cases, we found deposition of particular light chain constant region (immunoglobulin light chain constant region 2: IGLC2) at amyloid tissues. Materials and Methods: Amyloid tissues were either stained with conventional anti-human lambda LC rabbit antiserum (DAKO PO-0130) or rabbit anti-IGLC2 antibodies (made by SIGMA ALDLICH). Deposited peptide within amyloid lesion was examined using Congo Red staining followed by combination of laser micro-dissection and mass-spectrometry combining (LC-MS/MS). Plasma cells were purified with CD138-immunomagnetic beads from bone marrow aspirated obtained from two cases. Recombination of LC DNA in purified plasma cells was analyzed using PCR and direct nucleic acid sequencing. Results and Discussion: LC-MS/MS analysis revealed the presence of IGLC2 in all cases without showing positive signal by conventional antibody. Recombination of IGLC2 was detected in plasma cells. Deposition of IGLC2 was confirmed by IHC using anti-IGLC2 antibodies. The present data imply that “IGLC2-related amyloidosis” might be overlooked by conventional IHC unless utilizing mass-spectrometry. Utilization of antibodies for IGLC2 should contribute to improve quality of diagnosis for AL-amyloidosis. We consider that investigation of IGLC2 is important for not only making diagnosis but also analyzing pathogenesis of AL-amyloidosis.

Muscle-specific deletion of comparative gene identification-58 (CGI-58) causes muscle steatosis but improves insulin sensitivity in male mice.

Intramyocellular accumulation of lipids is often associated with insulin resistance. Deficiency of comparative gene identification-58 (CGI-58) causes cytosolic deposition of triglyceride (TG)-rich lipid droplets in most cell types, including muscle due to defective TG hydrolysis. It was unclear, however, whether CGI-58 deficiency-induced lipid accumulation in muscle influences insulin sensitivity. Here we show that muscle-specific CGI-58 knockout mice relative to their controls have increased glucose tolerance and insulin sensitivity on a Western-type high-fat diet, despite TG accumulation in both heart and oxidative skeletal muscle and cholesterol deposition in heart. Although the intracardiomyocellular lipid deposition results in cardiac ventricular fibrosis and systolic dysfunction, muscle-specific CGI-58 knockout mice show increased glucose uptake in heart and soleus muscle, improved insulin signaling in insulin-sensitive tissues, and reduced plasma concentrations of glucose, insulin, and cholesterol. Hepatic contents of TG and cholesterol are also decreased in these animals. Cardiac steatosis is attributable, at least in part, to decreases in cardiac TG hydrolase activity and peroxisome proliferator-activated receptor-α/peroxisome proliferator-activated receptor-γ coactivator-1-dependent mitochondrial fatty acid oxidation. In conclusion, muscle CGI-58 deficiency causes cardiac dysfunction and fat deposition in oxidative muscles but induces a series of favorable metabolic changes in mice fed a high-fat diet.

Maternal nicotine exposure during gestation and lactation induces cardiac remodeling in rat offspring.

The aim of this study was to evaluate the effects of maternal nicotine exposure on heart morphology and fibrosis in rat offspring. Nicotine was administered to pregnant Sprague-Dawley rats by using a subcutaneous osmotic mini-pump at a dose of 6 mg/kg/day from Gestational Days 7-21 or Gestational Day 7 to Postnatal Day 14. A control group received an equal volume of saline by the same route as nicotine. Rats born to prenatal nicotine-treated dams exhibited significantly greater cell width of cardiomyocytes, fewer cardiomyocyte nuclei number, higher β-myosin heavy chain and transforming growth factor (TGF-β1) expression, and higher collagen deposition in heart compared with rats born to normal saline-treated dams on Postnatal Days 7 and 21. Postnatal nicotine exposure further enhances these effects. We conclude that TGF-β1 may be involved in the pathogenesis of cardiac remodeling induced by maternal nicotine exposure during gestation and lactation in rat offspring.

Increased collagen deposition in the heart of chronically hypoxic ovine fetuses.

This study determined the effect of chronic intrauterine hypoxia on collagen deposition in the fetal sheep heart. Moderate or severe hypoxia was induced by placental embolization in chronically catheterized fetal sheep for 15 days starting at gestational day 116 ± 2 (term ∼147 days). The fetal right and left ventricle were evaluated for collagen content using a Sirius red dye and for changes in signaling components of pathways involved in collagen synthesis and remodeling using quantitative polymerase chain reaction and Western blot. In severely hypoxic fetuses (n = 6), there was a two-fold increase (P < 0.05) in the percentage staining for collagen in the right ventricle, compared with control (n = 6), whereas collagen content was not altered in the moderate group (n = 4). Procollagen I and III mRNA levels were increased in the right ventricle, two-fold (P < 0.05) and three-fold (P < 0.05), respectively, in the severe group relative to control. These changes were paralleled by a two-fold increase (P < 0.05) in mRNA levels of the pro-fibrotic cytokine, transforming growth factor β (TGF-β1), in the right ventricle. In the right ventricle, the mRNA levels of matrix metalloproteinase 2 (MMP-2) and its activator, membrane-type MMP (MTI-MMP) were increased five-fold (P = 0.06) and three-fold (P < 0.05), respectively, relative to control. Protein levels of TGF-β were increased in the left ventricle (P < 0.05). Thus, up-regulated collagen synthesis leading to increased collagen content occurs in the chronically hypoxic fetal heart and may contribute to the right ventricular diastolic and systolic dysfunction reported in human intrauterine growth restriction fetuses.

Effects of Oligomeric Grape Seed Proanthocyanidins on Heart, Aorta, Kidney in DOCA‐Salt Mice: Role of Oxidative Stress

Growing experimental and clinical data highlights the important roles of increased reactive oxygen species production in cardiovascular remodeling (CR). Oligomeric grape seed proanthocyanidins (GSPs) have been shown to be potent antioxidants and free radical scavengers. Mice were treated with DOCA-salt to induce CR and were given distilled water or oligomeric GSPs for 4 weeks. The heart weight (HW) index and kidney weight (KW) index were expressed as heart weight/body weight (HW/BW) and kidney weight/body weight (KW/BW); the histological changes were investigated by hematoxylin and eosin and Van Gieson staining.The endothelial-dependent vasodilation function induced by acetylcholine was investigated in isolated thoracic aorta ring. Colorimetric analysis was used to assay superoxide dismutase (SOD) activity, malondialdehyde (MDA) content and nitric oxide (NO) content in serum and hydroxyproline content in cardiac tissue. Administration of GSPs markedly alleviated the elevation of HW/BW ratio, KW/BW ratio and cross-sectional area of cardiomyocytes, decreased collagen deposition in heart and attenuated histopathology injury, and improves endothelial-dependent aorta ring relaxation in vitro accompany by increasing of NO content in serum. Meanwhile, treatment with GSPs significantly ameliorated oxidative stress via increasing SOD activities and decreasing MDA formation. These findings suggest that administration of GSPs has the potential to attenuate DOCA-salt induced CR and KH and preserve NO activity and endothelial function, which mechanism may contribute to its antioxidant characteristic, at least in part.

Reduction of thrombus size in murine models of thrombosis following administration of recombinant α1-proteinase inhibitor mutant proteins

The variant serpin α1-PI M358R inhibits thrombin and other proteases such as activated protein C (APC) and factor XIa. We previously described recombinant proteins HAPI M358R (α1-PI M358R containing an N-terminal extension corresponding to residues 1-75 of heparin cofactor II) and HAPI RCL5 (HAPI M358R with F352-I356 and I360 substituted for the corresponding residues of antithrombin), with enhanced selectivity for thrombin over APC inhibition. We tested the hypotheses that these recombinant proteins would limit thrombosis in three mouse models, and that the HAPI chimeric proteins would be more effective than α1-PI M358R. Recombinant serpins were purified from Escherichia coli by nickel chelate and ion exchange affinity chromatography, and administered to mice intravenously. HAPI RCL5 reduced incorporation of radiolabelled fibrin(ogen) into thrombi in the ferric chloride-injured vena cava in a dose-dependent manner; HAPI M358R was less effective and α1-PI M358R was without effect. In a model of murine endotoxaemia, HAPI RCL5 was more effective than α1-PI M358R in reducing radiolabelled fibrin(ogen) deposition in heart and kidneys; immunohistochemistry of tissue sections showed lesser staining with anti-fibrin(ogen) antibodies with both treatments. In the ferric chloride-injured murine carotid artery, administration of both recombinant serpins was equally effective in lengthening the vessel's time to occlusion. Our results show that the antithrombotic efficacy of the recombinant serpins correlates with their potency as thrombin inhibitors, since HAPI RCL5 inhibits thrombin, but not factors Xa, XIa, XIIa, or neutrophil elastase, more rapidly than α1-PI M358R.

Olive Leaf Extract Attenuates Cardiac, Hepatic, and Metabolic Changes in High Carbohydrate–, High Fat–Fed Rats1–3

Olive oil, an important component of the Mediterranean diet, produces cardioprotective effects, probably due to both oleic acid and the polyphenols such as oleuropein and hydroxytyrosol. Our aim in this study was to assess whether a polyphenol-enriched extract from the leaves of Olea europaea L. with oleuropein as the major component attenuated the cardiovascular, hepatic, and metabolic signs of a high-carbohydrate, high-fat (HCHF) diet (carbohydrate, 52%; fat, 24%, 25% fructose in drinking water) in rats. Male Wistar rats were fed either a cornstarch diet (CS) or a HCHF diet for a total of 16 wk. Diets of the treatment groups [CS+olive leaf extract (OLE) and HCHF+OLE] were supplemented with 3% OLE after 8 wk of being fed their respective CS or HCHF diets for a further 8 wk. After 16 wk, HCHF rats developed signs of metabolic syndrome, including elevated abdominal and hepatic fat deposition, collagen deposition in heart and liver, cardiac stiffness, and oxidative stress markers (plasma malondialdehyde and uric acid concentrations), with diminished aortic ring reactivity, abnormal plasma lipid profile, impaired glucose tolerance, and hypertension. Compared with HCHF rats, those in the HCHF+OLE group had improved or normalized cardiovascular, hepatic, and metabolic signs with the exception of elevated blood pressure. These results strongly suggest that an OLE containing polyphenols such as oleuropein and hydroxytyrosol reverses the chronic inflammation and oxidative stress that induces the cardiovascular, hepatic, and metabolic symptoms in this rat model of diet-induced obesity and diabetes without changing blood pressure.

The influence of different levels of dietary selenium on its distribution in the organs of broiler chickens

The content of selenium in basic broiler diets was 50 ? 10, 100 ? 10, 150 ? 10 and 250 ? 10 ?gkg-1. At day 14, 28 and 42 tissue samples were collected. Dynamics and selenium deposits in blood, liver, muscle and heart were estimated in relation to the level of dietary selenium and age of broilers. When broilers were fed with diet containing 250 ?gkg-1 Se, at day 42 the highest concentration of Se was in liver (570.8 ? 44.1 ?gkg-1), blood (430.3 ? 46.5 ?gL-1) and heart (237.5 ? 42.8 ?gkg-1). At day 14, increase of dietary selenium content from 50 ?gkg-1 to 250 ?gkg-1 was followed by decrease of selenium deposits in heart: when broilers were fed with basic diet containing 50 ?gkg-1 Se measured content of Se in heart muscle was 49.8 ? 15.6 ?gkg-1 (99.6%) while in broilers fed with basic diet containing 250 ?gkg-1 Se measured content of Se in heart was 147.2 ? 33.4 ?gkg-1 (58.9%), respectively. The point of saturation as well as maximal concentration of selenium in liver was reached in period between fourth (556.7 ? 40.6 ?gkg-1) and sixth (570.8 ? 44.1 ?gkg-1) week of age. Also, feeding with basic diet containing 250 ?gkg-1 Se the selenium blood level reached 162.8 ? 28.9 ?gL-1 already at day 14 that represent 65.1% of dietary selenium.