Metabolomics of repetitive myocardial stunning in chronic multivessel coronary artery stenosis: Effect of non-selective and selective ß1-receptor blockers.

Chronic coronary artery stenosis can lead to regional myocardial dysfunction in the absence of myocardial infarction by repetitive stunning, hibernation or both. The molecular mechanisms underlying repetitive stunning-associated myocardial dysfunction are not clear. We used non-targeted metabolomics to elucidate responses to chronically stunned myocardium in a canine model with and without ß-adrenergic blockade treatment. After development of left ventricular systolic dysfunction induced by ameroid constrictors on the coronary arteries, animals were randomized to 3 months of placebo, metoprolol or carvedilol. We compared these two ß-blockers with their different ß-adrenergic selectivities on myocardial function, perfusion and metabolic pathways involved in tissue undergoing chronic stunning. Control animals underwent sham surgery. Dysfunction in stunned myocardium was associated with reduced fatty acid oxidation and enhanced ketogenic amino acid metabolism, together with alterations in mitochondrial membrane phospholipid composition. These changes were consistent with impaired mitochondrial function and were linked to reduced nitric oxide and peroxisome proliferator-activated receptor signalling, resulting in a decline in adenosine monophosphate-activated protein kinase. Mitochondrial changes were ameliorated by carvedilol more than metoprolol, and improvement was linked to nitric oxide and possibly hydrogen sulphide signalling. In summary, repetitive myocardial stunning commonly seen in chronic multivessel coronary artery disease is associated with adverse metabolic remodelling linked to mitochondrial dysfunction and specific signalling pathways. These changes are reversed by ß-blockers, with the non-selective inhibitor having a more favourable impact. This is the first investigation to demonstrate that ß-blockade-associated improvement of ventricular function in chronic myocardial stunning is associated with restoration of mitochondrial function. KEY POINTS: The mechanisms responsible for the metabolic changes associated with repetitive myocardial stunning seen in chronic multivessel coronary artery disease have not been fully investigated. In a canine model of repetitive myocardial stunning, we showed that carvedilol, a non-selective ß-receptor blocker, ameliorated adverse metabolic remodelling compared to metoprolol, a selective ß1-receptor blocker, by improving nitric oxide synthase and adenosine monophosphate protein kinase function, enhancing calcium/calmodulin-dependent protein kinase, probably increasing hydrogen sulphide, and suppressing cyclic-adenosine monophosphate signalling. Mitochondrial fatty acid oxidation alterations were ameliorated by carvedilol to a larger extent than metoprolol; this improvement was linked to nitric oxide and possibly hydrogen sulphide signalling. Both ß-blockers improved the cardiac energy imbalance by reducing metabolites in ketogenic amino acid and nucleotide metabolism. These results elucidated why metabolic remodelling with carvedilol is preferable to metoprolol when treating chronic ischaemic left ventricular systolic dysfunction caused by repetitive myocardial stunning.

Proteolytic degradation of atrial sarcomere proteins underlies contractile defects in atrial fibrillation.

Atrial fibrillation (AFib) is the most common cardiac rhythm disturbance, often treated via electrical cardioversion. Following rhythm restoration, a period of depressed mechanical function known as atrial stunning occurs, suggesting that defects in contractility occur in AFib and are revealed upon restoration of rhythm. This project aims to define the contractile remodeling that occurs in AFib. To assess contractile function, we used a canine atrial tachypacing model of induced AFib. Mass spectrometry analysis showed dysregulation of contractile proteins in samples from AFib compared with sinus rhythm atria. Atrial cardiomyocytes show reduced force of contraction, decreased resting tension, and increased calcium sensitivity in skinned single cardiomyocyte studies. These alterations correlated with degradation of myofilament proteins including myosin heavy chain altering force of contraction, titin altering resting tension, and troponin I altering calcium sensitivity. We measured degradation of other myofilament proteins, including cardiac myosin binding protein C and actinin, that show degradation products in the AFib samples that are absent in the sinus rhythm atria. Many of the degradation products appeared as discrete cleavage products that are generated by calpain proteolysis. We assessed calpain activity and found it to be significantly increased. These results provide an understanding of the contractile remodeling that occurs in AFib and provide insight into the molecular explanation for atrial stunning and the increased risk of atrial thrombus and stroke in AFib.NEW & NOTEWORTHY Atrial fibrillation is the most common cardiac rhythm disorder, and remodeling during atrial fibrillation is highly variable between patients. This study has defined the biophysical changes in contractility that occur in atrial fibrillation along with identifying potential molecular mechanisms that may drive this remodeling. This includes proteolysis of several myofilament proteins including titin, troponin I, myosin heavy chain, myosin binding protein C, and actinin, which is consistent with the observed contractile deficits.

Decoding halal and jhatka slaughter: novel insights into welfare and protein biomarkers in slow-growing broiler chicken.

BACKGROUND: The first evidence of blood biochemical and proteomic changes in slow-growing broiler chicken subjected to ritual slaughter like halal (HS) and jhatka (JS) without stunning and commercial slaughter with electrical stunning (ES) was decoded. RESULTS: Significant stress indicators like cortisol and triiodothyronine were markedly elevated in JS birds, whereas increased (P < 0.05) levels of lactate dehydrogenase and creatine kinase were observed in JS and ES birds. Two-dimensional gel electrophoresis coupled to MALDI-TOF MS elucidated the overabundance of glyceraldehyde-3-phosphate dehydrogenase and l-lactate dehydrogenase that are positively correlated with stress in JS broilers. Bioinformatic analysis explored the multifaceted landscape of molecular functions. CONCLUSION: The study has uncovered that ritual slaughter performed without stunning against commercial slaughter with ES practices elicit varying levels of stress as evident from blood biochemistry and novel protein markers. © 2024 Society of Chemical Industry.

Comparison of recovery of sheep, goats, and calves from reversible electrical head-only and head-to-body stunning for halal meat production.

AIMS: To compare the recovery of lambs, goats, and calves from head-only (HO) or high-frequency head-to-body stunning and evaluate the complementary use of behaviour and electroencephalography (EEG) to assess return to consciousness after electrical stunning in these species. METHODS: Six-month-old lambs, adult goats and calves (< 7 days old) were subjected to reversible head-only stunning (50 Hz, 1 A, 2 seconds) or reversible high-frequency head-to-body stunning (RHTB: HO followed by 2,000 Hz, 2 A, 4-second stun to body). Following stunning, behavioural recovery was assessed in 21 lambs, 22 goats, and 20 calves. Latencies to first perform behaviours (end of convulsions, head lift, attempt to right, successful righting, attempt to stand, successful standing) after stunning were scored from video recordings. Recovery of electrical brain activity indicative of consciousness was assessed using EEG in a separate cohort of minimally-anaesthetised lambs, goats and calves (n = 20 per species). EEG traces collected before and after stunning were classified as normal, epileptiform, isoelectric, or transitional activity. Following stunning, the duration of epileptiform and isoelectric activity combined (states of brain activity incompatible with conscious awareness) was calculated, as was latency to return of normal (pre-stun) EEG. RESULTS: The RHTB stun was reversible in all three species, although one sheep failed to recover and was euthanised. Both methods caused tonic and clonic convulsions in all species. Behavioural recovery of sheep and calves was similar for both methods while goats took longer to recover from RHTB than HO stunning. There was no evidence of differences between methods in the duration of EEG incompatible with consciousness or the latency to recovery of normal EEG. CONCLUSIONS: Head-to-body stunning as applied here produced a reversible electrical stun in lambs, adult goats and young calves, although the benefits in terms of meat quality and operator safety are uncertain. Goats took longer to recover behaviourally from head-to-body stunning, possibly due to disrupted motor function, but there was no indication that post-stun unconsciousness lasted longer than following head-only stunning in any species. The normal behaviour for the animals' developmental age should be considered when deciding on behavioural indicators of recovery. The minimal anaesthesia model provided excellent quality EEG data that was valuable for interpretation of the behavioural responses. CLINICAL RELEVANCE: For the purposes of pre-slaughter stunning of sheep, goats and young calves, recovery appears comparable between the two methods, with all but 1/63 animals in the behaviour study recovering normal function.

Impact of stunning before slaughter on expression of skeletal muscles proteome in sheep.

The commercial production of halal and kosher meat and controversy surrounding the slaughter without stunning is rapidly growing across the globe. Huge global market for halal and kosher meat warrants conciliation of religious practices and animal welfare for the betterment of meat industry. In the present study, we investigated changes in muscle proteome of sheep (Ovis aries) subjected to either electrical stunning and slaughtering or slaughter without any stunning (halal). The 2DE gel analysis detected approximately 377 protein spots in which 243 (119 up regulated and 124 down regulated) protein spots were significantly (p ≤ 0.05) differentially expressed with a fold change ratio ≥1.5/≤1.5. The in-gel digestion and MALDI-TOF/TOF MS analysis of statistically significant protein spots revealed 35 differentially abundant proteins out of which 26 were up-regulated and 9 were down-regulated. The study demonstrated that slaughtering of sheep without stunning resulted in changes in the abundance of proteins involved in catalytic, structural, and stress related processes. This understanding of protein alterations in sheep slaughtered with and without stunning have the potential to act as possible signature for animal welfare index.

A review of the DTS: Diathermic Syncope® system with a discussion regarding its use for kosher slaughter (shechita).

Over the last several decades an alternative to current methods of stunning cattle has been developed. This system, DTS: Diathermic Syncope®, has been suggested to the Jewish and Muslim communities as a means to achieve pre-cut stunning in conformity with both religious and EU regulations without a need to resort to a derogation that permits an exemption from the EU requirement to pre-stun all animals undergoing slaughter. The developer's contention is that the system induces fainting, and thus should be acceptable to all groups, including the kosher (Jewish) and Halal (Muslim) consumer. A review of the system based on publications and reports from the developer itself suggests that in reality the system selectively heats the brain, leading to an epileptic-type seizure with tonic-clonic phases and unconsciousness lasting several minutes. It does not induce a (benign) faint, and use of the system might cause structural brain damage. Thus, this system is unlikely to be acceptable under Jewish religious law and its animal welfare value can be questioned.

The efficacy of electrical stunning of New Zealand rock lobster (Jasus edwardsii) and freshwater crayfish (Paranephrops zealandicus) using the Crustastun™.

Large numbers of decapod crustacea are farmed and harvested globally for human consumption. Growing evidence for the capacity of these animals to feel pain, and therefore to suffer, has led to increased concern for their welfare, including at slaughter. In New Zealand, decapod crustacea are protected by animal welfare legislation. There is a requirement that all farmed or commercially caught animals of these species killed for commercial purposes are first rendered insensible. The aim of this study was to evaluate the efficacy of the Crustastun™, a commercially available bench-top electrical stunner, in two commercially important New Zealand crustacean species; the rock lobster (Jasus edwardsii) and koura (freshwater crayfish [Paranephrops zealandicus]). Animals were anaesthetised via intramuscular injection of lidocaine and instrumented to record the electrical activity of the nervous system, prior to being stunned according to the manufacturer's instructions. Stunning efficacy was determined by analysing neural activity and observing behaviour post stunning. All ten P. zealandicus and three J. edwardsii appeared to be killed outright by the stun. Of the remaining J. edwardsii, six exhibited some degree of muscle tone and/or slow unco-ordinated movements of the limbs or mouthparts after stunning, although there was no recovery of spontaneous or evoked movements. One J. edwardsii was unable to be stunned successfully, likely due to its very large size (1.76 kg). None of the successfully stunned animals showed any evidence of return of awareness in the five minutes following stunning. It was concluded that the Crustastun™ is an acceptable method for killing P. zealandicus and for stunning all but the largest J. edwardsii.

Ginsenoside Rb1 Improves Post-Cardiac Arrest Myocardial Stunning and Cerebral Outcomes by Regulating the Keap1/Nrf2 Pathway.

The prognosis of cardiac arrest (CA) is dismal despite the ongoing progress in cardiopulmonary resuscitation (CPR). ginsenoside Rb1 (Gn-Rb1) has been verified to be cardioprotective in cardiac remodeling and cardiac ischemia/reperfusion (I/R) injury, but its role is less known in CA. After 15 min of potassium chloride-induced CA, male C57BL/6 mice were resuscitated. Gn-Rb1 was blindly randomized to mice after 20 s of CPR. We assessed the cardiac systolic function before CA and 3 h after CPR. Mortality rates, neurological outcome, mitochondrial homeostasis, and the levels of oxidative stress were evaluated. We found that Gn-Rb1 improved the long-term survival during the post-resuscitation period but did not affect the ROSC rate. Further mechanistic investigations revealed that Gn-Rb1 ameliorated CA/CPR-induced mitochondrial destabilization and oxidative stress, partially via the activation of Keap1/Nrf2 axis. Gn-Rb1 improved the neurological outcome after resuscitation partially by balancing the oxidative stress and suppressing apoptosis. In sum, Gn-Rb1 protects against post-CA myocardial stunning and cerebral outcomes via the induction of the Nrf2 signaling pathway, which may offer a new insight into therapeutic strategies for CA.

Stunning methods before slaughter induce oxidation changes of large yellow croaker during cold storage: the role of mitochondria and underlying mechanisms.

BACKGROUND: Improper stunning methods before slaughter could cause fish to deteriorate more quickly during cold storage. However, it is unclear how stunning methods affect the mitochondrial structure and the role of mitochondria in oxidation in muscle-based food. RESULTS: This study explored the potential mechanism of oxidation induced by different stunning methods (hit on the head, T1 ; gill cut, T2 ; immersion in ice/water slurry, T3 ; CO2 asphyxiated, T4 ; 40% CO2 + 30% N2 + 30% O2 , T5 ) in large yellow croaker during cold storage. The results showed that T4 samples had the minimum stress response and the mitochondrial membrane potential and permeability were less damaged. Besides, the mitochondrial functional structure and peroxisome of T4 samples were less damaged compared with other samples, which was reflected in higher total superoxide dismutase, catalase and glutathione peroxidase activities. In terms of oxidation indices, the T4 samples showed higher pH values and iron myoglobin contents and lower total volatile basic nitrogen and thiobarbituric acid reactive substances after 168 h cold storage, indicating that the T4 samples significantly maintained oxidative stability of large yellow croaker. CONCLUSION: The CO2 asphyxiation had the least oxidative damage to large yellow croaker during cold storage, possibly because it had the least effect on mitochondrial structure, reactive oxygen species and antioxidant enzyme activity. © 2023 Society of Chemical Industry.

Biomarker discovery and authentication of cold-slaughtered chicken through classical analytical procedures and mass spectrometry based proteomic approaches.

1. This study evaluated the suitability of routine analytical procedures and used mass spectrometry-based proteomic approaches to distinguish meat from dead chicken/ cold-slaughtered birds (CS), electrically stunned and slaughtered birds, as per standard protocols (ES), and birds slaughtered according to halal guidelines (HS).2. Meat from CS birds had lower (P < 0.05) pH, water-holding capacity and higher (P < 0.05) lipid oxidation, haem iron content, residual blood and total viable counts relative to ES and HS meat indicating poor quality.3. The results demonstrated the presence of unique protein bands on SDS-PAGE only in CS meat that can be used for routine screening.4. Protein analysis using MALDI-TOF mass spectrometry identified haemoglobin subunit alpha-A and alpha-D; Adenylate kinase isoenzyme 1 as reliable and stable marker proteins for authentication of dead chicken meat under raw and cooked conditions and halal slaughtered chicken, respectively.5. The methods used may be employed by the food safety and regulatory agencies for regular screening of meat quality and to authenticate CS or HS chicken.

Neurogenic stunned myocardium in acute ischemic stroke.

BACKGROUND AND PURPOSE: Neurogenic myocardial injury occurs as a result of dysregulation of autonomic nervous system. The aim of this study was to explore the frequency of elevated troponin and dynamic ST segment/T wave changes and their relation with left ventricular (LV) systolic functions in acute ischemic stroke patients. METHODS: One hundred and twenty-five patients (mean age: 65.1±15.2years, 76 male) presenting with acute ischemic stroke were consecutively included. 12-lead electrocardiogram was taken to assess dynamic ST segment/T wave changes, conventional transthoracic echocardiography to determine LV ejection fraction (LVEF). High-sensitive cardiac troponin I (hs-cTnI) level>0.04ng/mL was accepted as elevated. RESULTS: Twenty-seven patients (21.6%) had elevated hs-cTnI and 60 patients (48%) had dynamic ST segment/T wave changes. The stroke patients with elevated hs-cTnI had significantly higher NT-proBNP values (2302±3450pg/mL vs 799±2075pg/mL p<0.001) and higher frequency of ST segment/T wave changes (85.2% vs 37.8% p<0.001), and lower LVEF (52.2±13.6% vs 61.0±8.5% p=0.002) compared to patients with normal troponin levels. The patients with ST segment/T wave changes had significantly higher frequencies of hyper-lipidemia (31.7% vs 15.4% p=0.031) and coronary artery disease (CAD) (43.3% vs 13.8% p<0.001), hs-cTnI (0.19±0.55ng/mL vs 0.02±0.01ng/mL p<0.001) and NT-proBNP levels (1430±2564pg/mL vs 842±2425pg/mL p=0.016), and lower LVEF (56.1±11.7% vs 61.9±8.3% p=0.009). Linear regression analysis revealed presence of CAD, but not ST segment/T wave changes as an independent predictor of hs-cTnI (p=0.034). LVEF was independently associated with hs-cTnI (p=0.003) and presence of CAD (p=0.009) when adjusted by age, sex and presence of ST segment/T wave changes. CONCLUSION: Troponin elevation and ST segment/T wave changes occurring in patients suffering acute ischemic stroke, especially in those with CAD, may be a sign of neurogenic stunned myocardium.

Acute Heart Failure in Multisystem Inflammatory Syndrome in Children in the Context of Global SARS-CoV-2 Pandemic.

BACKGROUND: Cardiac injury and myocarditis have been described in adults with coronavirus disease 2019 (COVID-19). Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection in children is typically minimally symptomatic. We report a series of febrile pediatric patients with acute heart failure potentially associated with SARS-CoV-2 infection and the multisystem inflammatory syndrome in children as defined by the US Centers for Disease Control and Prevention. METHODS: Over a 2-month period, contemporary with the SARS-CoV-2 pandemic in France and Switzerland, we retrospectively collected clinical, biological, therapeutic, and early outcomes data in children who were admitted to pediatric intensive care units in 14 centers for cardiogenic shock, left ventricular dysfunction, and severe inflammatory state. RESULTS: Thirty-five children were identified and included in the study. Median age at admission was 10 years (range, 2-16 years). Comorbidities were present in 28%, including asthma and overweight. Gastrointestinal symptoms were prominent. Left ventricular ejection fraction was <30% in one-third; 80% required inotropic support with 28% treated with extracorporeal membrane oxygenation. Inflammation markers were suggestive of cytokine storm (interleukin-6 median, 135 pg/mL) and macrophage activation (D-dimer median, 5284 ng/mL). Mean BNP (B-type natriuretic peptide) was elevated (5743 pg/mL). Thirty-one of 35 patients (88%) tested positive for SARS-CoV-2 infection by polymerase chain reaction of nasopharyngeal swab or serology. All patients received intravenous immunoglobulin, with adjunctive steroid therapy used in one-third. Left ventricular function was restored in the 25 of 35 of those discharged from the intensive care unit. No patient died, and all patients treated with extracorporeal membrane oxygenation were successfully weaned. CONCLUSIONS: Children may experience an acute cardiac decompensation caused by severe inflammatory state after SARS-CoV-2 infection (multisystem inflammatory syndrome in children). Treatment with immunoglobulin appears to be associated with recovery of left ventricular systolic function.

Outcomes Among Patients With Left Ventricular Assist Devices Receiving Maintenance Outpatient Hemodialysis: A Case Series.

RATIONALE & OBJECTIVE: The incidence of left ventricular assist device (LVAD) implantation as destination therapy for heart failure is increasing and kidney failure requiring maintenance hemodialysis is a common complication. Because little is known about the safety or efficacy of outpatient hemodialysis among patients with LVADs, this study sought to describe their clinical course. STUDY DESIGN: Case series of patients with an LVAD undergoing maintenance outpatient hemodialysis whose clinical data were obtained from an electronic medical record. SETTING & PARTICIPANTS: Adults who received an LVAD, survived to hospital discharge, and were subsequently treated with maintenance hemodialysis by a not-for-profit dialysis provider between 2011 and 2019. RESULTS: 11 patients were included. 6 had a known history of chronic kidney disease. Patients underwent outpatient hemodialysis for a mean duration of 165.2 (range, 31-542) days, during which they were treated with 544 total dialysis sessions. 6 of these sessions were stopped early due to dialysis-related adverse events (1.1%). More than 80% of follow-up time was spent out of the hospital; however, 55% of patients were rehospitalized within 1 month of starting outpatient hemodialysis. The most common reason for hospitalization was infection (32%), followed by hypervolemia (14%), and cerebrovascular accident or transient ischemic attack (11%). 4 patients recovered kidney function, 1 underwent combined heart and kidney transplantation, 2 continued treatment, 2 died, and 2 were lost to follow-up. LIMITATIONS: Retrospective design, small number of cases, and lack of complete follow-up data. CONCLUSIONS: Approximately half the patients with complete follow-up either recovered kidney function or underwent combined heart and kidney transplantation. This case series demonstrates that outpatient hemodialysis centers, in partnership with LVAD treatment teams, can successfully provide hemodialysis to patients on LVAD support.

Aneurysmal subarachnoid hemorrhage as a trigger for Takotsubo syndrome: a comprehensive review.

Takotsubo syndrome (TTS) can result in acute heart failure and lead to a potentially life-threatening complication of aneurysmal subarachnoid hemorrhage (aSAH). The incidence of TTS in aSAH is less than 10% of all patients with aSAH, with a preponderance of postmenopausal women. Early indicators of TTS include elevated serum troponin levels and electrocardiographic abnormalities. The key finding is left ventricular wall motion abnormality. Echocardiography and coronary angiography help to establish the diagnosis. Vasopressors, milrinone, levosimendan, insulin, and anticoagulation may be required. The value of beta-blockers is a matter of controversy. TTS must not delay the treatment of a ruptured aneurysm. The clinical outcome in patients with aSAH and TTS is mostly determined by the aSAH and not the TTS.

[Prognostic implications of stone heart syndrome in cardiac arrest]. / Stellenwert des "Stone-heart"-Phänomens bei Herz-Kreislauf-Stillstand.

The stone heart syndrome is defined as an ischemic systolic contracture of the heart and also termed contractile cardiac arrest. It was first described in 1972 by the American cardiac surgeon Denton Cooley, who observed this phenomenon during bypass surgery. It is mostly the result of prolonged cardiac arrest where myocardial cells suffer hypoxia or anoxia. Insufficient forward blood flow and a decreased pressure gradient in the central aorta lead to reduced coronary perfusion. The resulting anaerobic metabolism causes an ischemic contracture as described in the stone heart syndrome. This article presents three cases of patients with traumatic cardiac arrest (TCA) and myocardial contracture in postmortem computed tomography (PMCT) and discuss the origins of the stone heart syndrome as well as its implications in cardiopulmonary resuscitation.

Quantitative proteomic and phosphoproteomic profiling of ischemic myocardial stunning in swine.

Despite decades of research on the pathophysiology of myocardial stunning, protein changes and/or phosphorylation status underlying alterations in cardiac function/structure remain inadequately understood. Here, we utilized comprehensive and quantitative proteomic and phosphoproteomic approaches to explore molecular mechanisms of myocardial stunning in swine. The closed-chest swine (n = 5 pigs) were subjected to a 10-min left anterior descending coronary artery (LAD) occlusion producing regional myocardial stunning. Tissues from the ischemic LAD region and a remote nonischemic area of the left ventricle were collected 1 h after reperfusion. Ion current-based proteomics (IonStar) and quantitative phosphoproteomics were employed in parallel to identify alterations in protein level and site-specific phosphorylation changes. A novel swine heart protein database exhibiting high accuracy and low redundancy was developed here to facilitate comprehensive study. Further informatic investigations identified potential protein-protein interactions in stunned myocardium. In total, we quantified 2,099 protein groups and 4,699 phosphorylation sites with only 0.4% missing values. Proteomic analyses revealed downregulation of contractile function and extracellular matrix remodeling. Meanwhile, alterations in phosphorylation linked with contractile dysfunction and apoptotic cell death were uncovered. NetworKIN/STRING analysis predicted regulatory kinases responsible for altered phosphosites, such as protein kinase C-mediated phosphorylation of cardiac troponin I-S199 and CaMKII-mediated phosphorylation of phospholamban-T17. In summary, the ion current-based proteomics and phosphoproteomics reliably identified novel alterations in protein content and phosphorylation contributing to contractile dysfunction, extracellular matrix (ECM) damage, and programmed cell death in stunned myocardium, which corroborate well with our physiological observations. Moreover, this work developed a comprehensive database of the swine heart proteome, a highly valuable resource for future translational research in porcine models with cardiovascular diseases.NEW & NOTEWORTHY We first used ion current-based proteomics and phosphoproteomics to reliably identify novel alterations in protein expression and phosphorylation contributing to contractile dysfunction, extracellular matrix (ECM) damage, and programmed cell death in stunned myocardium and developed a comprehensive swine heart-specific proteome database, which provides a valuable resource for future research in porcine models of cardiovascular diseases.

The punched-out tissue complex (skin-bone "imprimatum") in shots from captive-bolt guns: does it act as a secondary projectile?

From the first half of the twentieth century to the present day, injuries and fatalities from captive-bolt livestock stunners are a major topic in forensic medicine. The vast majority of cases account for suicides with the frontal, temporal, parietal, and occipital regions being the most common sites of entrance (in descending order of frequency). Due to the limited length of the bolt, the corresponding wound channel within the braincase is only several centimeters long. It has been a controversial subject for a long time, whether the skin-bone complex punched out by the conically grooved end of the steel rod may act as a "secondary projectile" being propelled beyond the actual path of the bolt. To answer this question, experimental shots from various types of captive bolt-guns were fired to simulants. Video-documentation employing a high-speed motion camera showed that the punched-out pieces of skin and bone did not move further than the bolt. Thus, a secondary extension of the total wound channel could not be observed. However, the suction effect caused by the bolt's rearward movement may induce a slight retrograde displacement of the skin-bone complex.

Better characterization of dipyridamole-induced myocardial stunning by systolic wall thickening. A gated perfusion SPECT study.

AIM AND PATIENTS: The aim of the present study was to assess the additional value of systolic wall thickening to myocardial perfusion in diagnosing myocardial stunning induced by dipyridamole infusion. We selected 52 ischemic patients (43 males; mean age 65.5 ± 7.64), with CAD documented by angiography. Ischemia was defined as a summed difference score ≥ 5. All patients underwent a 2-day gated perfusion SPECT protocol. The patients received a dose of 740 MBq of 99mTc-tetrofosmin after stress and at rest. RESULTS: The post-stress LVEF was significantly lower than rest LVEF (48.3% ± 14.5% vs. 50.7% ± 15%; P = 0.0001). The wall thickening summed difference score was 3.97 ± 3.84 (P = 0.0001). At a multivariate regression analysis, only WT-SDS as independent variable was significantly correlated with myocardial ischemia (SDS) (P = 0.001). We divided patients according to SDS in those with mild (SDS < 8) and severe (SDS ≥ 8) ischemia. WT-SDS, but not ∆LVEF, was significantly different between groups. CONCLUSIONS: WT-SDS showed a better correlation with the degree of ischemia than the depression in the global function of the left ventricle. It allowed to better identify the stunning phenomenon in patients submitted to pharmacological stress.

Is humane slaughtering of rainbow trout achieved in conventional production chains in Germany? Results of a pilot field and laboratory study.

BACKGROUND: Rainbow trout, Oncorhynchus mykiss, is an important fish in European freshwater aquaculture. This industry sector is dominated by small family-owned enterprises located in rural areas. A large percentage of rainbow trout produced by these small enterprises is marketed directly and killed on demand and not processed in commercial processing plants. EU and national regulations stipulate that fish shall be stunned prior to killing and slaughter. The overall objective of this study was to monitor how stunning interventions were integrated into the production chains of German conventional trout aquaculture in order to safeguard animal welfare during stunning and killing. For this, the stunning and slaughtering processes were monitored on 18 rainbow trout farms in various German federal states. During the on-farm research, (i) the stunning success, (ii) injuries related to the stunning procedure, (iii) duration between stunning and killing, and (iv) visible responses at the time of slaughtering were assessed as welfare indicators. In addition, haematological and biochemical blood parameters were measured as indicators for physiological stress. Due to the fact that stunning interventions should induce a loss of consciousness in fish, in a laboratory study, it was examined whether the absence of the brainstem/ behavioural responses, opercular movements (OM) or eye-rolling reflex (vestibulo-ocular reflex, VOR) was correlated with the stage of insensibility. RESULTS: The majority of rainbow trout farms applied manual percussion (38%) or electrical stunning (48%), while on 14% of the farms, the fish were stunned by electrical stunning which was immediately followed by manual percussion. After percussive stunning, about 92.3% of the rainbow trout displayed no OM or VOR as brainstem/ behavioural indicators of consciousness. This percentage varied on farms which applied electrical stunning. While on the majority of farms, 95 to 100% of the fish were unconscious according to the observation of brainstem/ behavioural indicators, the stunning intervention was less effective on farms where rainbow trout were stunned at current densities below 0.1 A dm2 or for a few seconds only. The laboratory study confirmed that the absence of brainstem/ behavioural indicators correlated with the absence of visually evoked responses (VER) of the brain to light stimuli as a neuronal indicator of insensibility. Therefore, the brainstem/ behavioural signs can be used to interpret the stage of insensibility in rainbow trout. A stage of insensibility could safely be induced by exposing portion-sized rainbow trout to an electric current density above 0.1 A dm2. This was not influenced by the orientation of the electric field. CONCLUSIONS: In conventional aquaculture, rainbow trout can effectively be stunned by manual percussion or electrical stunning. Consciousness can be monitored by the absence of opercular movements or the eye-rolling reflex, which are lost approximately at the same time as neurological responses like VER. For safeguarding animal welfare during stunning and killing of rainbow trout in conventional production processes, the stunning process requires careful attention and the operating personnel need to be trained in using the stunning devices and recognising indicators of consciousness.

No evidence of cardiac stunning or decoupling immediately after cardiopulmonary bypass for elective coronary surgery.

BACKGROUND: There is significant uncertainty regarding the timing of onset of cardiovascular stunning after cardiac surgery. Cardiovascular stunning is affecting both contractility (Ees) and arterial load. Arterial load may be represented by arterial elastance (Ea) and participates in ventriculo-arterial coupling through the Ea/Ees ratio, giving information on efficiency and performance. An alternative approach to ventriculo-arterial interaction is oscillatory power fraction (OPF). The aim of this study was to investigate the immediate beat-to-beat effects of on-pump coronary artery bypass graft (CABG) surgery on contractility, cardiac power parameters, arterial load and ventriculo-arterial coupling as well as classical haemodynamic parameters. METHODS: We included 41 patients scheduled for fast-track CABG surgery. Measurements were taken before and after cardiopulmonary bypass. A flow and pressure curve were recorded from transoesophageal pulsed wave Doppler and a radial artery catheter, respectively. This enabled the calculation of stroke work, total cardiac energy delivery, OPF and Ea/Ees ratio. Routine haemodynamic monitoring provided the classical haemodynamic parameters. RESULTS: Immediately after cardiopulmonary bypass there was no firm evidence for alterations in contractility, stroke work, stroke volume or arterial elastance. Ea/Ees ratio and OPF remained unchanged. CONCLUSIONS: There was no evidence for clinically relevant cardiac stunning or altered arterial load immediately after cardiopulmonary bypass for CABG surgery. The unchanged Ea/Ees ratio and OPF are indicating unchanged cardiac efficiency before and after cardiopulmonary bypass. This indicates that in elective CABG patients cardiovascular stunning is perhaps a phenomenon of inflammation and not immediate ischaemia-reperfusion injury or mechanical handling.