Cardiac Specimens Research Articles

Prophylactic Evidence of MSCs-Derived Exosomes in Doxorubicin/Trastuzumab-Induced Cardiotoxicity: Beyond Mechanistic Target of NRG-1/Erb Signaling Pathway

Trastuzumab (Trz) is a humanized monoclonal antibody targeting epidermal growth factor receptor 2 (HER2; ErbB2). The combined administration of Trz and doxorubicin (DOX) has shown potent anti-cancer efficacy; however, this regimen may be accompanied by severe cardiac toxicity. Mesenchymal stem cells (MSCs)-derived exosomes are nanosized vesicles that play a crucial role in cell–cell communication and have shown efficacy in the treatment of various diseases. In this study, we aim to investigate the cardioprotective effects of MSCs-derived exosomes in a DOX/Trz- mediated cardiotoxicity model, and the possible mechanisms underlying these effects are elucidated. Forty-nine male rats were randomly assigned into four groups: Group I (control); Group II (Dox/Trz); Group III (protective group); and Group IV (curative group). Cardiac hemodynamic parameters, serum markers of cardiac injury, oxidative stress indices, and cardiac histopathology were investigated. Further, transcript profile of specific cardiac tissue injury markers, apoptotic markers, and fibrotic markers were analyzed using qRT-PCR, while the protein expressions of pAkt/Akt, pERK/ERK, pJNK/JNK, pJNK/JNK, and pSTAT3/STAT3 were evaluated by ELISA. Additionally, cardiac mirR-21 and miR-26a were assessed. A combined administration of DOX/Trz disrupted redox and Ca2+ homeostasis in cardiac tissue induced myocardial fibrosis and myofibril loss and triggered cardiac DNA damage and apoptosis. This cardiotoxicity was accompanied by decreased NRG-1 mRNA expression, HER2 protein expression, and suppressed AKT and ERK phosphorylation, while triggering JNK phosphorylation. Histological and ultra-structural examination of cardiac specimens revealed features typical of cardiac tissue injury. Moreover, a significant decline in cardiac function was observed through biochemical testing of serum cardiac markers and echocardiography. In contrast, the intraperitoneal administration of MSCs-derived exosomes alleviated cardiac injury in both protective and curative protocols; however, superior effects were observed in the protective protocol. The results of the current study indicate the ability of MSCs-derived exosomes to protect from and attenuate DOX/Trz-induced cardiotoxicity. The NRG-1/HER2, MAPK, PI3K/AKT, PJNK/JNK, and PSTAT/STAT signaling pathways play roles in mediating these effects.

BRG1 is a biomarker of hypertrophic cardiomyopathy in human heart specimens

Hypertrophic cardiomyopathy (HCM) is a genetic disease of the sarcomere that causes otherwise unexplained cardiac hypertrophy and is associated with sudden death. While previous studies showed the role of the epigenetic modifier Brg1 in mouse models of HCM, additional work is needed to identify its role in humans. We tested the hypothesis that BRG1 expression is increased in periods of cardiac remodeling during fetal growth and in development of HCM. We employed immunohistochemical staining to evaluate protein expression of BRG1 in 796 human cardiac specimens (81 from patients with HCM) and describe elevated BRG1 expression in human fetal hearts in early development. In addition, we not only demonstrate increased expression of BRG1 in HCM, but we also show that other diseases that lead to heart failure have similar BRG1 expression to healthy controls. Inhibition of BRG1 in human induced pluripotent stem cell-derived cardiomyocytes significantly decreases MYH7 and increases MYH6, suggesting a regulatory role for BRG1 in the pathological imbalance of the two myosin heavy chain isoforms in human HCM. These data are the first demonstration of BRG1 as a specific biomarker for human HCM and provide foundation for future studies of epigenetics in human cardiac disease.

Assaying fibroblast activation protein (FAP) expression in vivo and in vitro for possible targeting with chimeric antigen receptor (CAR) T cells

Fibroblast activation protein (FAP) is a marker of activated fibroblasts in various human cardiovascular diseases. FAP is of special interest because it is expressed exclusively in activated, pathogenic fibroblasts and has been proposed as a target for chimeric antigen receptor (CAR) T cell ablation therapy. It has been shown previously that fibroblasts from patients with hypertrophic and dilated cardiomyopathy express FAP, while fibroblasts from non‐failing hearts did not. The aim of the project was to examine FAP expression in other human cardiac diseases by obtaining heart samples from patients who suffered diseases characterized by cardiac fibrosis so that we could determine which cardiac diseases might be amenable to FAP CAR T therapy. In addition, we sought to characterize an in vitro model of FAP expression to aid in the ability to assess the effectiveness of engineered T cells to kill FAP‐expressing cells. Heart tissue from patients diagnosed with various cardiac disorders, including idiopathic giant cell myocarditis, COVID‐19, and sarcoidosis were stained for FAP expression and compared to non‐failing, control hearts. The results showed significant FAP expression in all cardiac specimens with interstitial fibrosis compared to non‐failing hearts. Notably, samples from patients who died from COVID‐19 exhibited significant fibrosis and FAP expression. This suggests that there may be a large and growing population that may be amenable to anti‐fibrotic therapy. In order to develop an in vitro model of FAP expression to assay anti‐FAP CAR T cell activity, we treated IMR‐90 human fibroblasts with TGF‐B, a known activator of fibrosis, and assayed for FAP expression. TGF‐B exposure resulted in an increase in FAP expression 24‐hours after treatment. Our observation of fibroblast restricted FAP expression in multiple different cardiovascular diseases supports the potential use of FAP CAR T cell therapy as an anti‐fibrotic treatment in multiple heart diseases.

The utility of a structured, interactive cardiac anatomy teaching session for resident education.

Paediatric residents are often taught cardiac anatomy with two-dimensional images of heart specimens, or via imaging such as echocardiography or computed tomography. This study aimed to determine if the use of a structured, interactive, teaching session using heart specimens with CHD would be effective in teaching the concepts of cardiac anatomy. The interest amongst paediatric residents of a cardiac anatomy session using heart specimens was assessed initially by circulating a survey. Next, four major cardiac lesions were identified to be of interest: atrial septal defect, ventricular septal defect, tetralogy of Fallot, and transposition. A list of key structures and anatomic concepts for these lesions was developed, and appropriate specimens demonstrating these features were identified by a cardiac morphologist. A structured, interactive, teaching session was then held with the paediatric residents using the cardiac specimens. The same 10-question assessment was administered at the beginning and end of the session. The initial survey demonstrated that all the paediatric residents had an interest in a cardiac anatomy teaching session. A total of 24 participated in the 2-hour session. The median pre-test score was 45%, compared to a median post-test score of 90% (p < 0.01). All paediatric residents who completed a post-session survey indicated that the session was a good use of educational time and contributed to increasing their knowledge base. They expressed great interest in future sessions. A 2-hour hands-on cardiac anatomy teaching session using cardiac specimens can successfully highlight key anatomic concepts for paediatric residents.

Histopathological Evaluation of Cardiac Lesions in Autopsy Specimens

ABSTRACT context: Evaluation of cardiac autopsy specimens is crucial for identifying concealed cardiovascular pathology and providing insights into the pathogenesis and progression of disease. Hence, this study was undertaken to evaluate the histopathological features of cardiac lesions in autopsy specimens. method: The was a prospective cross-sectional study conducted on cardiac specimens from routine autopsies over a period of three and half years, between January 2018 and June 2021, at M.S. Ramaiah Medical College and Hospitals, Bengaluru. The hearts were evaluated grossly and microscopically to identify the various histopathological changes. results: A total of 250 cases, with mean age of 39±15 years and male-female ratio of 4.4:1, were evaluated. In 207 (82.8%) cases 299 morphological lesions were identified, comprised of coronary atherosclerosis (52.4%), myocardial infarction (26%), aortic atherosclerosis (21.2%), cardiac hypertrophy (12%), valvular disease (2%), aortic dissection (2%), cardiac tamponade (1.6%), pericarditis (1.2%), myocarditis (0.8%) and cardiomyopathy (0.4%). conclusion: Cardiac lesions were detected in 82.8% in autopsies. The most frequent finding is coronary atherosclerosis with triple vessel involvement being the commonest. Severe luminal occlusion, calcification and thrombosis are most frequent in Left anterior descending coronary artery. The next most common lesion is Myocardial Infarction, the most significant pathogenetic mechanism of which is coronary atherosclerosis. There is a strong association between coronary and ascending aorta atherosclerosis. Thus, many preventable and concealed lesions are discovered at cardiac autopsies and this data is important for assessing disease trends and introducing new interventions and therapeutic management regimes

Cardiac microcalcifications in transthyretin (ATTR) amyloidosis

BackgroundBone tracers bind to amyloid-containing heart of most patients with ATTR amyloidosis. Amyloid deposits outside the heart are often scarce and bone scintigraphy is increasingly often used to diagnose cardiac involvement. However, the nature of the binding of bone tracers to the heart is not clear. ObjectiveTo identify possible calcium deposits in hearts with amyloid, explaining bone tracer binding. Methods and resultsFormalin-fixed and paraffin embedded cardiac specimens from three patients with ATTR and one with AL amyloidosis, all with cardiac deposits, were studied. The specimens covered large parts of the heart. Sections were stained immunohistochemically for ATTR deposits and according to von Kóssa for calcifications. The study identified in all hearts, but particularly in the ATTR materials, focal, tight swarms of tiny calcifications. These were sometimes associated with amyloid but found as frequent in areas without such deposits. Autoradiography with [99mTc]Tc labelled 3,3-disphos-phono-1,2-propanodicarboxylic acid (DPD) revealed labelling in von Kóssa positive areas. Electron microscopically the particles were not amorphous but had a complex structured appearance and were often surrounded by a membrane, indicating a cellular origin. Labelling with antibodies against ubiquitin and P62 pointed to result from autophagy. ConclusionsOur study indicates that binding of skeletal probes to amyloid-containing hearts depends on an irregular presence of clouds of very tiny calcifications, which seem not to be directly associated with amyloid fibrils. Therefore, [99mTc]Tc-DPD bone scans can be considered surrogate markers of ATTR amyloid but have to be used carefully to estimate amyloid amount or disease progression.

Congenital heart defects in the fetus and embryological classification: Cladistics and phylogeny

Congenital heart defects (CHD) are a common cause of fetal death and termination of pregnancy for fetal anomaly (TPFA). No series with sufficient cohort, based on description of the anatomic phenotype of CHD discovered in fetal period, have been published until now. To phenotype all the fetal cardiac specimens of the M3 C collection, which counts more than 1500 hearts. A complete morphological examination of each specimen according to segmental analysis was performed by two observers (MH, LH). We determined the main CHD according to the 11 categories and 23 subcategories of the clinical and anatomical classification of CHD and we coded the associated lesions with IPCCC codes. These codes were recorded in a database including photographs for each heart. To date, we have analyzed 758 cardiac specimens. Among them, 77 were normal (10.2%). The most frequent groups of CHD were: anomalies of the ventricular outflow tracts (VOT, 34%), functionally univentricular hearts (FUV, 24%) and anomalies of atrioventricular junctions and valves (AAVJV, 15%). As an example of a phylogenetic analysis starting from a specific anomaly, a right aortic arch was most frequent in double outlet right ventricle with subaortic ventricular septal defect (VSD) (39%), tetralogy of Fallot group (24%) and outlet VSD (9%). It was almost absent in other anomalies of VOT and other CHD ( Fig. 1 ). These first results confirm the predominance of the anomalies of the VOT. The high rate of FUV and AAVJV underlines the selection bias related to the high number of TPFA for severe CHD or chromosomal abnormality. This anatomical phenotyping of the collection should allow us to identify rare associations of malformations and could change our current way of grouping some phenotypes together. This could help us, by integrating current embryological knowledge, to elaborate a cladistics analysis of CHD, starting from one part of the phenotype.

Perfusion-distention fixation of heart specimens: a key step in immortalizing heart specimens for wax infiltration and generating 3D imaging data sets for reconstruction and printed 3D models

Perfusion-distention fixation by gravity is a fundamental cardiac predissection technique that may be used to preserve the 3D anatomy of a cardiac specimen. The equipment and steps are illustrated, and examples of cardiac specimens preserved in various methods including wet specimens, wax-infiltrated hearts, and 3D-printed models, created after performing this technique, are shown.

Abstract 12950: PUMA Sequestration by IP3Rs Underlies Maladaptive Cardiac Fibrosis

Introduction: Following an ischemic injury, the persistence of cardiac myofibroblast (myo-FBs) contributes to fibrosis and adverse myocardial remodeling, especially if myoFBs remain active in otherwise healthy areas of the heart, away from the site of injury. The exact mechanisms underlying myo-FB activation and persistence remain unknown. Hypothesis: Since recent genome-wide association (GWA) studies have identified a significant association between gain-of-function mutations in genes encoding for inositol 1,4,5-trisphosphate receptors (IP3Rs) and ischemic heart disease, we hypothesized that IP3Rs could be involved in post-ischemic cardiac fibrosis. Methods: We applied an established model of ischemia/reperfusion (I/R) to a newly generated triple KO mouse specifically lacking all isoforms of IP3Rs in cardiac myo-FBs, in order to avoid compensatory responses. Results: After I/R, IP3Rs level is not changed in the scar area, whereas it is markedly upregulated in the remote regions. Compared to control littermates, KO mice display a reduced interstitial fibrosis and an attenuated myocardial dysfunction following I/R. Mechanistically, we show for the first time that IP3Rs modulate apoptosis in myo-FBs following ischemic injury in a calcium-independent manner, i.e. by directly sequestrating the pro-apoptotic factor PUMA (p53 upregulated modulator of apoptosis), thereby favoring the detrimental myoFBs persistence in the remote areas. Strikingly, these results were confirmed in human cardiac specimens from patients with heart failure, as well as in vitro in human FBs, in which IP3Rs are shown to regulate cell proliferation and migration. Conclusions: Our findings indicate that the IP3R-PUMA interaction selectively regulate the activation of cardiac myo-FBs located in the regions remote from the ischemic injury.

Cardiogenic shock complicating multisystem inflammatory syndrome following COVID-19 infection: a case report

BackgroundWith the high prevalence of COVID-19 infections worldwide, the multisystem inflammatory syndrome in adults (MIS-A) is becoming an increasingly recognized entity. This syndrome presents in patients several weeks after infection with COVID-19 and is associated with thrombosis, elevated inflammatory markers, hemodynamic compromise and cardiac dysfunction. Treatment is often with steroids and intravenous immunoglobulin (IVIg). The pathologic basis of myocardial injury in MIS-A, however, is not well characterized. In our case report, we obtained endomyocardial biopsy that revealed a pattern of myocardial injury similar to that found in COVID-19 cardiac specimens.Case presentationA 26-year-old male presented with fevers, chills, headache, nausea, vomiting, and diarrhea 5 weeks after his COVID-19 infection. His SARS-CoV-2 PCR was negative and IgG was positive, consistent with prior infection. He was found to be in cardiogenic shock with biventricular failure, requiring inotropes and diuretics. Given concern for acute fulminant myocarditis, an endomyocardial biopsy (EMB) was performed, showing an inflammatory infiltrate consisting predominantly of interstitial macrophages with scant T lymphocytes. The histologic pattern was similar to that of cardiac specimens from COVID-19 patients, helping rule out myocarditis as the prevailing diagnosis. His case was complicated by persistent hypoxemia, and a computed tomography scan revealed pulmonary emboli. He received IVIg, steroids, and anticoagulation with rapid recovery of biventricular function.ConclusionsMIS-A should be considered as the diagnosis in patients presenting several weeks after COVID-19 infection with severe inflammation and multi-organ involvement. In our case, EMB facilitated identification of MIS-A and guided therapy. The patient’s biventricular function recovered with IVIg and steroids.

Aortic stenosis: correlation of prenatal echocardiography to postmortem histology

Abstract Background Foetal aortic stenosis (AS) is a challenging congenital heart disease considering its potential to progress during the course of pregnancy. Especially at midgestation, it remains extremely difficult to distinguish the cases that end up biventricular from the cases that will develop into an hypoplastic left heart syndrome. Purpose To test the hypothesis that the degree of myocardial maturation is a possible predictor of biventricular outcome, we present 4 cases of foetal AS with a varying degree of severity and uniquely correlate differences in myocardial function based on prenatal echocardiography to their post-mortem histopathologic maturation. Methods We selected 4 cases with midgestational AS from our tertiary foetal cardiology service between 2018–2020. Speckle tracking recordings of the cardiac four-chamber view were performed during routine foetal echocardiography to quantify myocardial wall motion as a marker for myocardial function. Three cases decided to terminate the pregnancy and donated the cardiac specimen. Immunohistochemical labelling (ICH) against key markers for myocardial maturation (troponin-I, N-cadherin, connexin-43, MLC2A, MLC2V and α-SMA) and fibrosis (Sirius Red) were compared with 2 normal foetal cardiac specimens. Results Two cases with critical AS presented extremely decreased global and segmental longitudinal strain (GLS and SLS) values (GLS −2% and −0.9%) in the left ventricle (LV), indicating an impaired myocardial wall deformation. Post-mortem ICH showed overt endocardial fibro-elastosis (EFE) and pathological fibrosis patterns in the subendocardial layer which was remarkably spatially correlated to the EFE. The cardiomyocytes were disorganised with reduced expression of troponin-I and disturbed expression of connexin-43. The remaining 2 cases had normal LV appearance on foetal echocardiography, showing a mild reduction in left ventricular GLS and SLS (GLS −11.8% and −14.2%). Post-mortem ICH of 1 of these cases showed mild EFE with a milder fibrosis pattern. Cardiomyocytes were less disorganised but also showed a disturbed expression of connexin-43. The 4th case continued the pregnancy and had a biventricular outcome. Conclusions This is a unique case series showing that myocardial function correlates with high extent to histology. The degree of the reduction in myocardial function corresponded with the amount of pathological fibrosis patterns and disorganisation of the cardiomyocyte network. Myocardial wall motion on foetal echocardiography seems to hold promise as a possible marker for cardiac maturation. Funding Acknowledgement Type of funding sources: None. Speckle tracking and fibrosis patterns

Deficient Myocardial Organization and Pathological Fibrosis in Fetal Aortic Stenosis-Association of Prenatal Ultrasound with Postmortem Histology.

In fetal aortic stenosis (AS), it remains challenging to predict left ventricular development over the course of pregnancy. Myocardial organization, differentiation and fibrosis could be potential biomarkers relevant for biventricular outcome. We present four cases of fetal AS with varying degrees of severity and associate myocardial deformation on fetal ultrasound with postmortem histopathological characteristics. During routine fetal echocardiography, speckle tracking recordings of the cardiac four-chamber view were performed to assess myocardial strain as parameter for myocardial deformation. After pregnancy termination, postmortem cardiac specimens were examined using immunohistochemical labeling (IHC) of key markers for myocardial organization, differentiation and fibrosis and compared to normal fetal hearts. Two cases with critical AS presented extremely decreased left ventricular (LV) strain on fetal ultrasound. IHC showed overt endocardial fibro-elastosis, which correlated with pathological fibrosis patterns in the myocardium and extremely disturbed cardiomyocyte organization. The LV in severe AS showed mildly reduced myocardial strain and less severe disorganization of the cardiomyocytes. In conclusion, the degree of reduction in myocardial deformation corresponded with high extent to the amount of pathological fibrosis patterns and cardiomyocyte disorganization. Myocardial deformation on fetal ultrasound seems to hold promise as a potential biomarker for left ventricular structural damage in AS.

Detection of Periodontal Pathogens in Oral Samples and Cardiac Specimens in Patients Undergoing Aortic Valve Replacement: A Pilot Study.

This observational study aimed to: (i) assess the presence of periodontal disease among patients requiring aortic valve replacement; (ii) investigate the presence of oral pathogens in aortic valve specimens and compare them with the microorganisms detected in the oral cavity. Twenty-six patients (15 men and 11 women) were scheduled to be visited the day before the cardiac surgery: periodontal conditions were accurately registered through clinical and radiographic examinations; dental plaque or salivary samples were collected. Valve specimens were collected during surgical aortic valve replacement and analyzed for pathogens detection through microbiological 16SrRna gene sequencing. Bacteria found in plaque samples and valve specimens were assessed according to oral and periodontal conditions. A qualitative comparison between oral and cardiac profiles of the microorganisms detected was performed. The overall number of patients examined for soft tissues conditions was 19, as 7 patients were edentulous. Twelve and three individuals, respectively, presented moderate and severe periodontitis. Nine valves were found to be positive for the presence of oral and periodontopathic bacterial DNA. The microbial species found in valve samples of patients with periodontitis suggest that the presence of these microorganisms in valvular tissue seems to be not coincidental.

Congenital heart defects in the foetus and embryological classification: Cladistic et phylogeny

Congenital heart defects (CHD) are a common cause of fetal death and termination of pregnancy for fetal anomaly (TPFA). No series with sufficient cohort, based on description of the anatomic phenotype of CHD discovered in fetal period, have been published until now. The aim of our study is to phenotype all the fetal cardiac specimens of the M3 C collection, which counts more than one thousand hearts. A complete morphological examination of each specimen according to segmental analysis was performed by two observers (MH,LH). We determined the main CHD according to the 11 categories et 23 sub-categories of the clinical and anatomical classification of CHD [2] and we coded the associated lesions with IPCCC codes according to the ICD-11 CHD classification [1] . These codes were recorded in a database created for that purpose including photographs for each heart. To date, we have analyzed 758 cardiac specimens. Among them, 77 were normal hearts (10,2%). The most frequent main groups of CHD were: anomalies of the ventricular outflow tracts (VOT) (231, 34%), functionally univentricular hearts (FUV) (165, 24%) and anomalies of atrioventricular junctions and valves (AVJV) (106, 15%). The most frequent associated lesions were: anomalies of the valves (all valve type; 409, 60%), ventricular hypoplasia (265, 39%), ventricular septal defects (233, 34%), interatrial communications (204, 30%), anomalies of the arterial duct (152, 22%). These first Results confirm the predominance of the anomalies of the VOT. The high rate of FUV and anomalies of AVJV underlines the selection bias related to the high number of TPFA for severe CHD or chromosomal abnormality. This anatomical phenotyping of the collection should allow us to identify rare associations of malformations and could change our current way of grouping some phenotypes together. This could help us, by integrating current embryological knowledge, to elaborate a cladistic analysis of CHD.

SARS-CoV-2 Myocarditis: Insights Into Incidence, Prognosis, and Therapeutic Implications.

Purpose of ReviewIn coronavirus disease 2019 (COVID-19), myocardial injury occurs frequently in severe or critically ill hospitalized patients, yet myocarditis is much less common. In this context, revisiting the definition of myocarditis is appropriate with a specific focus on diagnostic and management considerations in patients infected with SARS-CoV-2.Recent FindingsPathologic cardiac specimens from patients with COVID-19 suggest a mixed inflammatory response involving lymphocytes and macrophages, and importantly, cellular injury occurs predominantly at the level of pericytes and endothelial cells, less often involving direct myocyte necrosis. In COVID-19, the diagnosis of myocarditis has understandably been based predominantly on clinical criteria, and the number of patients with clinically suspected myocarditis who would meet diagnostic histological criteria is unclear. Echocardiography and cardiac magnetic resonance are important diagnostic tools, although the prognostic implications of abnormalities are still being defined. Importantly, SARS-CoV2 myocarditis should be diagnosed within an appropriate clinical context and should not be based on isolated imaging findings. Therapies in COVID-19 have focused on the major clinical manifestation of pneumonia, but the promotion of viral clearance early in the disease could prevent the development of myocarditis, and further study of immunosuppressive therapies once myocarditis has developed are indicated.SummaryA strict and uniform approach is needed to diagnose myocarditis due to SARS-CoV-2 to better understand the natural history of this disease and to facilitate evaluation of potential therapeutic interventions. A methodological approach will also better inform the incidence of COVID-19 associated myocarditis and potential long-term health effects.

The Possible Role of Allicin in Ameliorating Azithromycin Induced Cardiotoxicity in Adult Male Albino Rat: A Histological and Immunohistochemical Study

Abstract Background: Azithromycin (AZ) is a broad-spectrum macrolide that is incorporated in the treatment of various infectious diseases, and recently enlisted in the protocol of Covid-19 management. Allicin is a classical garlic extraction with cardioprotective, antioxidant, anti-inflammatory, and anti-apoptotic properties.Aim: To evaluate the cardiotoxic effect of AZ and evaluate the possible protective effect of Allicin against AZ cardiotoxicity using various histological and immunohistochemical techniques.Materials and Methods: Forty adult male albino rats were randomly divided into four main groups: group-I acted as a control group, group-II was given Allicin (20 mg/kg/day) orally for consecutive 14 days , group-III was given AZ (30 mg/kg/day) orally for consecutive 14 days and group-IV was given both AZ and Allicin in the same doses for consecutive 14 days. The cardiac specimens were processed for different histological and imunohistochemical techniques. Morphometrical and statistical studies were also done. Results: Azithromycin induced several myocardial changes in the form of focal areas of destruction of cardiac muscle fibers, darkly stained pyknotic nuclei and cytoplasmic vacuoles. Wavy muscle fibers, widening of the intercellular spaces, mononuclear cellular infiltration, fatty infiltration and hemorrhage were obvious. Dilated and congested blood vessels were also noticed. A significant increase in the mean area percentage of both Masson and α-SMA immunoreactivity was detected while a significant decrease in the mean area percentage of Bcl-2 was demonstrated. In contrast, most of the histological changes disappeared by Allicin co-treatment except in few localized areas. Conclusion: Azithromycin induced several destructive changes in cardiac muscle fibers. Allicin had a potent ameliorative role in prevention of cardiotoxicity induced by AZ

Effects of platelet rich plasma on experimentally induced diabetic heart injury

Diabetic heart is one of the common complications of diabetes mellitus. Platelet-rich plasma (PRP) is an autologous product rich in growth factors that can enhance tissue regeneration. This work was conducted to study the PRP ability to improve diabetes-inducing cardiac changes. Also, it sheds more light on the possible mechanisms through which PRP induces its effects. Rats were divided into; control, PRP, diabetic, and PRP-diabetic groups. Cardiac specimens were obtained and processed for biochemical, histological, and immunohistochemical study. The diabetic group exhibited a significant increase in cardiac oxidative stress, inflammation, and cardiac injury markers if compared with the control group. Additionally, the cardiac tissue showed variable morphological changes in the form of focal distortion and loss of cardiac myocytes. Distorted mitochondria and heterochromatic nuclei were observed in the cardiac muscle fibers. The mean number of charcoal-stained macrophages, and mean area fraction for collagen fibers, mean number of PCNA-immune positive cardiac muscle were significantly decrease in PRP- diabetic group. Collectively, the results showed that PRP treatment ameliorated most of all these previous changes. ConclusionPRP ameliorated the diabetic cardiac injury via inhibition of oxidative stress and inflammation. It was confirmed by biochemical, histological, and immunohistochemical study. It could be concluded that PRP could be used as a potential therapy for diabetic heart.

Congenital heart defects in the foetus and embryological classification: Cladistic et phylogeny

Congenital heart defects (CHD) are a common cause of fetal death and termination of pregnancy for fetal anomaly (TPFA). No series with sufficient cohort, based on description of the anatomic phenotype of CHD discovered in fetal period, have been published until now. The aim of our study is to phenotype all the fetal cardiac specimens of the M3C collection, which counts more than one thousand hearts. A complete morphological examination of each specimen according to segmental analysis was performed by two observers (MH, LH). We determined the main CHD according to the 11 categories and 23 sub-categories of the clinical and anatomical classification of CHD and we coded the associated lesions with IPCCC codes according to the ICD-11 CHD classification. These codes were recorded in a database created for that purpose including photographs of each heart. To date, we have analyzed 697 cardiac specimens. Among them, 64 were normal hearts (9%). The most frequent main groups of CHD were: anomalies of the ventricular outflow tracts (VOT) (197, 28%), functionally univentricular hearts (FUV) (164, 24%), anomalies of atrioventricular junction and valves (AVJV) (95, 14%). The most frequent associated lesions were: anomalies of the valves (all valve type; 442, 63%), ventricular septal defects (241, 35%), interatrial communications (198, 28%), ventricular hypoplasia (172, 25%), anomalies of the arterial duct (152, 22%). These first results confirm the predominance of the anomalies of the VOT. The high rate of FUV and anomalies of AVJV underlines the selection bias related to the high number of TPFA for severe CHD or chromosomal abnormality. This anatomical phenotyping of the collection should allow us to identify rare associations of malformations and could change our current way of grouping some phenotypes together. This could help us, by integrating current embryological knowledge, to elaborate a cladistic analysis of CHD.

Lactoferrin Ameliorates Azithromycin-induced Cardiac Injury: Insight into Oxidative Stress/TLR4 /NF-κB Pathway

Introduction: Azithromycin, a widely used antibacterial agent, is also considered the cornerstone of the management protocols for COVID-19 infection, particularly in Egypt, due to its antiviral impact and preventive effects against secondary bacterial infections. However, even at pharmacological doses, azithromycin develops rapid cardiac toxicity. Objectives: Our study here aims at investigating the cardioprotective potentials of lactoferrin, a natural compound used primarily for boosting intestinal iron absorption, to counteract azithromycin-induced cardiac toxicity in experimental rats when administered concurrently. Methods: Male Sprague Dawley rats received either lactoferrin (200 mg/kg/day, orally) or saline for 10 days. Induction of cardiac toxicity was initiated on the 6th day via oral administration of azithromycin (20mg/kg/day, orally) for 5 successive days, concurrently with either lactoferrin or vehicle. Cardiac injuries were confirmed via Electrocardiogram (ECG) recording, assessment of cardiac function biomarkers, assessment of cardiac expression of nuclear factor erythroid 2–related factor 2 (Nrf2), interleukin-1β (IL-1β) and IL-10, measurement of cardiac oxidant/antioxidant balance, immunohistochemical staining against toll-like receptor-4 (TLR4), and nuclear factor-κB (NF-κB), and histopathological examination of HE-stained cardiac specimens. Results: Administration of azithromycin denoted a significant cardiac tissue injury as demonstrated by upregulated serum levels of cardiac biomarkers, namely total creatine Kinase (CK), Creatine kinase-MB (CK-MB), lactate dehydrogenase (LDH), and Alkaline phosphatase (ALP). Furthermore, it upregulated cardiac content of malondialdehyde (MDA) and nitric oxide (NO) and cardiac inflammatory markers, such as TLR4, NF-κB, and IL-1β. Similarly, azithromycin-downregulated reduced glutathione (GSH) cardiac content and the anti-inflammatory protective indicators, IL-10 and Nrf2. Additionally, azithromycin-induced cardiotoxicity was evinced by ECG pattern and histopathologic deterioration. Oral Lactoferrin showed cardioprotective potentials and counteracted the azithromycin-induced cardiac toxicity, downregulating cardiac biomarkers. Moreover, it restored cardiac oxidant/antioxidant balance, decreased cardiac TLR4, NF-κB, and IL-1β expression, and increased IL-10 and Nrf2 expression. Lactoferrin Cardioprotective potentials were also evinced by enhanced ECG pattern and histopathological examination. Conclusion: Finally, we support the use of lactoferrin with azithromycin intake that induced cardiac toxicity.

Cardiac ankyrin repeat protein contributes to dilated cardiomyopathy and heart failure.

Cardiac ankyrin repeat protein (CARP) is a cardiac-specific stress-response protein which exerts diverse effects to modulate cardiac remodeling in response to pathological stimuli. We examined the role of CARP in postnatal cardiac development and function under basal conditions in mice. Transgenic mice that selectively overexpressed CARP in heart (CARP Tg) exhibited dilated cardiac chambers, impaired heart function, and cardiac fibrosis as assessed by echocardiography and histological staining. Furthermore, the mice had a shorter lifespan and reduced survival rate in response to ischemic acute myocardial infarction. Immunofluorescence demonstrated the overexpressed CARP protein was predominantly accumulated in the nuclei of cardiomyocytes. Microarray analysis revealed that the nuclear localization of CARP was associated with the suppression of calcium-handling proteins. In vitro experiments revealed that CARP overexpression resulted in decreased cell contraction and calcium transient. In post-mortem cardiac specimens from patients with dilated cardiomyopathy and end-stage heart failure, CARP was significantly increased. Taken together, our data identified CARP as a crucial contributor in dilated cardiomyopathy and heart failure which was associated with its regulation of calcium-handling proteins.