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Objectives Methane (CH4) breath test is an established diagnostic method for gastrointestinal functional disorders. Our aim was to explore the possible link between splanchnic circulatory changes and exhaled CH4 in an attempt to recognize intestinal perfusion failure. Design Randomized, controlled in vivo animal study. Setting University research laboratory. Subjects Anesthetized, ventilated Sprague-Dawley rats (280 ± 30 g) and Vietnamese minipigs (31 ± 7 kg). Interventions In the first series, CH4 was administered intraluminally into the ileum before 45 minutes mesenteric ischemia or before reperfusion in non-CH4 producer rats to test the appearance of the gas in the exhaled air. In the porcine experiments, the superior mesenteric artery was gradually obstructed during consecutive, 30-minute flow reductions and 30-minute reperfusions achieving complete occlusion after four cycles (n = 6), or nonocclusive mesenteric ischemia was induced by pericardial tamponade (n = 12), which decreased superior mesenteric artery flow from 351 ± 55 to 182 ± 67 mL/min and mean arterial pressure from 96.7 ± 18.2 to 41.5 ± 4.6 mm Hg for 60 minutes. Measurements and main results Macrohemodynamics were monitored continuously; RBC velocity of the ileal serosa or mucosa was recorded by intravital videomicroscopy. The concentration of exhaled CH4 was measured online simultaneously with high-sensitivity photoacoustic spectroscopy. The intestinal flow changes during the occlusion-reperfusion phases were accompanied by parallel changes in breath CH4 output. Also in cardiac tamponade-induced nonocclusive intestinal ischemia, the superior mesenteric artery flow and RBC velocity correlated significantly with parallel changes in CH4 concentration in the exhaled air (Pearson's r = 0.669 or r = 0.632, respectively). Conclusions we report a combination of in vivo experimental data on a close association of an exhaled endogenous gas with acute mesenteric macro- and microvascular flow changes. Breath CH4 analysis may offer a noninvasive approach to follow the status of the splanchnic circulation.

OBJECTIVES Extracorporeal circulation induces cellular and humoral inflammatory reactions, thus possibly leading to detrimental secondary inflammatory responses. Previous data have demonstrated the bioactive potential of methane and confirmed its anti-inflammatory effects in model experiments. Our goal was to investigate the in vivo consequences of exogenous methane administration on extracorporeal circulation-induced inflammation. METHODS Two groups of anaesthetized Vietnamese minipigs (non-treated and methane treated, n = 5 each) were included. Standard central cannulation was performed, and extracorporeal circulation was maintained for 120 min without cardiac arrest or ischaemia, followed by an additional 120-min observation period with haemodynamic monitoring. In the methane-treated group, 2.5% v/v methane–normoxic air mixture was added to the oxygenator sweep gas. Blood samples through the central venous line and tissue biopsies from the heart, ileum and kidney were taken at the end point to determine the whole blood superoxide production (chemiluminometry) and the activity of xanthine-oxidoreductase and myeloperoxidase, with substrate-specific reactions. RESULTS Methane treatment resulted in significantly higher renal blood flow during the extracorporeal circulation period compared to the non-treated group (63.9 ± 16.4 vs 29.0 ± 9.3 ml/min). Whole blood superoxide production (548 ± 179 vs 1283 ± 193 Relative Light Unit (RLU)), ileal myeloperoxidase (2.23 ± 0.2 vs 3.26 ± 0.6 mU/(mg protein)) and cardiac (1.5 ± 0.6 vs 4.7 ± 2.5 pmol/min/mg), ileal (2.2 ± 0.6 vs 7.0 ± 3.4 pmol/min/mg) and renal (1.2 ± 0.8 vs 13.3 ± 8.0 pmol/min/mg) xanthine-oxidoreductase activity were significantly lower in the treated group. CONCLUSIONS The addition of bioactive gases, such as methane, through the oxygenator of the extracorporeal circuit represents a novel strategy to influence the inflammatory effects of extracorporeal perfusion in cardiac surgical procedures.

Our aim was to characterize the main components of the nitrosative response with quantitative changes of the nitrergic myenteric neurons in adjacent intestinal segments after transient superior mesenteric artery occlusion. We also tested the hypothesis that exogenous methane may modulate the evolution of nitroxidation by influencing xanthine oxidoreductase (XOR) activity. The microcirculatory consequences of a 50 min ischemia or ischemia-reperfusion were investigated in anesthetized rats (n = 124) inhaling normoxic air with or without 2.2% methane. XOR activities, nitrogen monoxide (NO), nitrite/nitrate (NOx), and nitrotyrosine levels were measured, together with relative nitrergic neuron ratios from duodenum, ileum and colon samples. The effects of methane on XOR were also examined in vitro. The intramural flow stopped only in the ileum during ischemia. The highest baseline XOR activity was found in the duodenum, which increased further during ischemia. NO and nitrotyrosine levels rose, and the nNOS-immunopositive neuron ratio and NOx level both dropped. Reperfusion uniformly elevated XOR activity and nitrotyrosine formation, with the highest level attained in the duodenum, where the nitrergic neuron ratio remained depressed. These alterations were eliminated in methane-treated animals, XOR activity and nitrotyrosine formation decreased in all sites, and the duodenal nitrergic neuron ratio was re-established. The inhibitory effect of methane on XOR-linked nitrate reductase activity was also demonstrated in vitro. With segment-specific microcirculatory alterations, the risk for nitrosative stress is highest in transiently hypoxic tissues with high endogenous XOR activities. The XOR-inhibitory effect of methane can reduce nitroxidation and protects the nitrergic neuron population in such conditions.

Absztrakt: A pericardialis tamponad a szivsebeszeti ellatas egyik fontos surgőssegi korkepe, amelynek hattereben szamos koroki tenyező allhat. A pericardialis folyadekfelszaporodas diagnosztikaja es terapiaja a klinikai gyakorlatban ma mar sikeres, ugyanakkor a kialakulo kardiogen sokk kovetkezmenyeinek eredmenyes kezelese meg sok esetben kihivast jelent. Az akut pericardialis tamponad es a kronikus pericardialis folyadekgyulem hatekony kezelesi tervehez a korelettani folyamat megertese szukseges. A jelen kozlemeny celja, hogy a pericardialis tamponad jelensegeről es jelentősegeről is osszefoglalo kepet nyujtson, az orvostorteneti hatter es a fontosabb korelettani tenyezők ismertetesen at egeszen az allatkiserletes modellalkotas szerepeig. Orv Hetil. 2018; 159(5): 163–167.

Absztrakt: Bevezetés: A pericardialis tamponád az életet közvetlenül veszélyeztető, alacsony perctérfogatú kórállapot, amelynek keringési következményei minden szövet, köztük a vese és a bélnyálkahártya mikrokeringését is súlyosan érintik. Célkitűzés: Kísérletünk célja a pericardialis tamponád klinikai képének modellezésére alkalmas, hemodinamikailag stabil, kontrollálható állatmodell kifejlesztése volt. Módszerek: Altatott vietnami törpesertések két csoportjában laparotomiát követően a szívburokba a rekeszen keresztül, thoracotomia nélkül kanült helyeztünk. Az első, álműtött csoport kontrollként szolgált (n = 6), a második csoportban a pericardium saját vérrel való feltöltésével 60 perces tamponádot hoztunk létre (n = 6). Monitoroztuk a tamponád és az azt követő 180 perces reperfúzió alatt a makrohemodinamikát és a vékonybél mikrohemodinamikai változásait. Vizsgáltuk a fehérvérsejt-akkumulációt jelző mieloperoxidáz (MPO) -enzimaktivitást és pásztázó lézeres endomikroszkóppal a vékonybél in vivo szövettani elváltozásait. Eredmények: A tamponád következtében megemelkedett a centrális vénás nyomás és a szívfrekvencia, lecsökkent az artériás középnyomás, az arteria mesenterica superior (355,5 ± 112,4 vs. 182,0 ± 59,1 ml/min) és az arteria renalis (159,63 ± 50,7 vs. 35,902 ± 27,9 ml/min) áramlása, valamint az ileum mikrokeringése. Az MPO-aktivitás szignifikánsan megemelkedett (3,66 ± 1,6 vs. 7,01 ± 1,44 mU/mg fehérje) és romlott a vékonybél in vivo szövettani képe. Következtetések: Kísérleti modellünk alkalmas a szívtamponád jellegzetes keringési és biokémiai következményeinek vizsgálatára, reprodukálható, klinikailag releváns állatmodellnek tekinthető.

Absztrakt: Bevezetés: A nem okkluzív mesenterialis ischaemia (NOMI) anatómiai okok nélkül alakul ki, korai diagnosztizálása nehéz, terápiája sem megoldott. Modellkísérleteinkben a komplement C5a gátlásán keresztül a komplementaktiváció szerepét vizsgáltuk a kórfolyamatban. Anyagokés módszerek: A NOMI rövid és hosszú távú hemodinamikai és gyulladásos következményeinek jellemzéséhez Sprague–Dawley patkányokban (n = 28) 60 perces parciális aortaokklúziót (PAO; hasi aorta, truncus coeliacustól proximálisan; arteria femoralis középnyomása: 30–40 Hgmm), míg vietnami törpesertésekben (n = 19) 60 perces pericardialis tamponádot (artériás középnyomás: 40–50 Hgmm) hoztunk létre. A PAO, valamint a tamponád 45. percében komplement C5a-gátló kezelést alkalmaztunk (acetil-peptid-A; 4 mg/kg iv.). Az állatokban monitoroztuk a makro- és mikrokeringést, mértük a leukocytainfiltrációt, a gyulladásos mediátorok (endothelin, HMGB-1) plazmaszintjét. Eredmények: Patkányokban a PAO megszüntetése után 24 órával a C5a gátlása csökkentette a szisztémás gyulladásos válasz részeként megemelkedett perctérfogatot (203,1 ± 5 vs. 269,6 ± 8,1 ml/‌min/‌kg) és arteria mesenterica superior (AMS) áramlását, fokozta az ileum mucosa mikrokeringését (833,5 ± 33,8 vs. 441,9 ± 22,4 μm/s). Az acetil-peptid-A sertésekben átmenetileg növelte az AMS áramlását és tartósan az ileummucosa-kapillárisáramlást (648,1 ± 45,4 vs. 329,8 ± 12,6 μm/s). A kezelt állatoknál alacsonyabb gyulladásos mediátorszintet és szöveti leukocytainfiltrációt mértünk mindkét NOMI-modellben. Következtetések: A komplementaktiváció jelentős szerepet játszik a NOMI alatt bekövetkező makro- és mikrokeringési zavar kialakulásában, a C5a gátlása a gyulladásos folyamat mérséklése mellett befolyásolja a NOMI hemodinamikai következményeit is.

We have developed a novel compound from acetylsalicylic acid (ASA) and 2-amino-2-(hydroxymethyl)-1,3-propanediol (Tris) precursors with ASA-like anti-inflammatory efficacy and reduced the mucosa-damaging side-effects. Our aim was to examine local and remote consequences of ASA-Tris administration in 2-,4-,6-trinitrobenzene-sulfonic acid (TNBS)-induced colitis as compared to ASA or mesalamine (5-aminosalicylate) treatment. Sprague–Dawley rats were randomized to five groups (n = 6, each), and TNBS enemas were performed. Group 1 was the negative control; group 2 was the untreated colitis group. 12 hour after colitis induction repeated doses of ASA, ASA-Tris (both 0.55 mmol/kg) and mesalamine (0.77 mmol/kg) were given 3 times daily for 3 days to groups 3–5. On day 3 of colitis, the in vivo histology of the colon and stomach was investigated. Tissue xanthine-oxidoreductase, myeloperoxidase, nitrite/nitrate changes, and circulating TNF-alpha levels were measured. In addition, liver mitochondria were examined with high-resolution respirometry to analyze alterations in the electron transport chain. TNBS enema significantly elevated inflammatory enzyme activities, NO production, TNF-alpha concentration, and induced morphological damage in the colon. ASA-treatment reduced the inflammatory marker levels and mucosal injury in the colon, but gastric tissue damage was present. ASA-Tris- and mesalamine-treatments significantly reduced the cytokine levels, inflammatory enzyme activities, and colonic mucosal damage without inducing gastric injury. Also, ASA significantly reduced the Complex IV-linked respiration of liver mitochondria, which was not observed after ASA-Tris-treatment. As compared to ASA, ASA-Tris conjugation provides significant protection against the colonic injury and cytokine-mediated progression of inflammatory events in experimental colitis without influencing the gastric epithelial structure.

Background Nonocclusive mesenteric ischemia (NOMI) can evolve in a variety of low-flow states. Although the mechanisms leading to NOMI-related intestinal necrosis are largely unknown, circumstantial evidence suggests that excessive vasoconstriction and complement activation both play important roles in this process. Because targeting of the circulatory malfunction of the splanchnic area could be of therapeutic relevance, we set out to investigate the long-term effects of treatment with a complement C5a antagonist in a rat model of partial aortic occlusion (PAO)-induced transient mesenteric hypoperfusion. Methods The mean arterial pressure of the splanchnic area was kept between 30 and 40 mm Hg by 60 minutes of PAO in anesthetized male Sprague-Dawley rats. C5a inhibitor acetyl-peptide-A (AcPepA; 4 mg kg−1 intravenously) or vehicle administration was initiated at the 45th minute of PAO. After 24 hours, the animals were reanesthetized to record the macrohemodynamics and ileal microcirculation, and plasma and tissue samples were taken for determination of high-mobility group box protein-1 (HMGB-1), endothelin-1, tumor necrosis factor (TNF)-α levels, and small intestinal leukocyte infiltration. Epithelial structural changes were visualized by in vivo confocal laser scanning endomicroscopy. Results At 24 hours after PAO, mean arterial pressure, heart rate, and cardiac output were significantly greater, the intestinal intramural microcirculation was significantly impaired, and plasma HMGB-1, endothelin-1, TNF-α levels, the degree of epithelial damage and leukocyte infiltration was increased. The AcPepA treatment moderated the hemodynamic and microcirculatory changes, and decreased inflammatory activation and histologic signs of mucosal damage. Conclusion C5a inhibition ameliorated the potentially harmful local mesenteric hypoperfusion and global long-term inflammatory consequences of PAO. This approach is of promise for use in NOMI-associated situations.

Acute pericardial tamponade is one of the most emergent clinical scenarios in cardiac surgery. With numerous causes in the background, pericardial tamponade can lead to cardiogenic shock and death. In modern diagnostic era, the recognition of pericardial tamponade is simple, but its management and the long-term effects can still be challenging. Without the detailed understanding of the pathophysiological pathways diagnostic and therapeutic management plans of pericardial tamponade is very difficult. The aim of this review was to give a complex picture of pericardial tamponade, from its role in medical history to pathophysiology and its significance in surgical experimental models. Orv Hetil. 2018; 159(5): 163-167.

Pericardial tamponade (PT) is a life-threatening condition, with low cardiac output. The hemodynamic consequences of PT can severely affect the circulation of all tissues, including the microcirculation of the kidneys and the intestinal mucosa. Our aim was to develop a hemodynamically stable and controllable large animal model of PT to study the consequences of cardiogenic shock.Two groups of anesthetized vietnamese minipigs (n = 6, both groups) were used. Following laparotomy, a cannula was fixed into the pericardium through the diaphragm without thoracotomy. A sham-operated group served as control, in the second group 60-min PT was induced by intrapericardial injection of heparinised own blood. Throughout PT and 180-min reperfusion, macrohemodynamics, renal circulation and mesenteric macro- and microcirculation were monitored. Myeloperoxidase (MPO) activity was measured and in vivo histology was performed by confocal laser scanning endomicroscopy.The PT increased central venous pressure, heart rate and decreased mean arterial pressure, mesenteric flow (from 355.5 ± 112.4 vs 182.0 ± 59.1 ml/min) and renal arterial flow (from 159.63 ± 50.7 vs 35.902 ± 27.9 ml/min) and the microcirculation of the ileum. Elevated MPO activity (3.66 ± 1.6 vs 7.01 ± 1.44 mU/mg protein) and injury of the ileal mucosa were present also.The reproducible large animal model is suitable for clinically relevant investigations of the hemodynamic and biochemical consequences of PT.

Non-occlusive mesenteric ischemia (NOMI) develops without anatomical causes. Early diagnosis is challenging and treatments are of questionable effectiveness. We investigated the role of complement activation in the pathophysiology of NOMI in animal models through the inhibition of complement C5a.60-min partial aortic occlusion (PAO; abdominal aorta, proximal to celiac trunk; mean arterial pressure: 30-40 mmHg) was established in Sprague-Dawley rats (n = 28) and 60-min cardiac tamponade in minipigs (n = 19; mean arterial pressure: 40-50 mmHg) to observe short- and long-term circulatory and inflammatory consequences of NOMI. Macro- and microhemodynamics, leukocyte infiltration, plasma levels of inflammatory mediators (endothelin, HMGB-1) were measured. C5a inhibitor (Acetyl-Peptid-A; 4 mg/kg iv) was administered at the 45th min of PAO or tamponade, respectively.Twenty-four hours after PAO systemic inflammatory response increased cardiac output and superior mesenteric artery flow (SMAF). C5a inhibition reduced the elevated cardiac output (203.1 ± 5 vs 269.6 ± 8.1 ml/min/kg) and SMAF and increased ileal microcirculation (833.5 ± 33.8 vs 441.9 ± 22.4 μm/s). In pigs, after the tamponade, C5a inhibition reduced the immediate hemodynamic disturbances, temporarily increased SMAF and permanently the ileal microcirculation. The Acetyl-Peptid-A treatment reduced leukocyte infiltration and plasma levels of inflammatory mediators in both NOMI models.Complement activation plays central role in the macro- and microcirculatory disturbance during NOMI. C5a inhibition reduces the inflammatory activation and influences the hemodynamic consequences of experimental NOMI.

Background Methane is part of the gaseous environment of the intestinal lumen. The purpose of this study was to elucidate the bioactivity of exogenous methane on the intestinal barrier function in an antigen-independent model of acute inflammation. Methods Anesthetized rats underwent sham operation or 45-min occlusion of the superior mesenteric artery. A normoxic methane (2.2%)-air mixture was inhaled for 15 min at the end of ischemia and at the beginning of a 60-min or 180-min reperfusion. The integrity of the epithelial barrier of the ileum was assessed by determining the lumen-to-blood clearance of fluorescent dextran, while microvascular permeability changes were detected by the Evans blue technique. Tissue levels of superoxide, nitrotyrosine, myeloperoxidase, and endothelin-1 were measured, the superficial mucosal damage was visualized and quantified, and the serosal microcirculation and mesenteric flow was recorded. Erythrocyte deformability and aggregation were tested in vitro. Results Reperfusion significantly increased epithelial permeability, worsened macro- and microcirculation, increased the production of proinflammatory mediators, and resulted in a rapid loss of the epithelium. Exogenous normoxic methane inhalation maintained the superficial mucosal structure, decreased epithelial permeability, and improved local microcirculation, with a decrease in reactive oxygen and nitrogen species generation. Both the deformability and aggregation of erythrocytes improved with incubation of methane. Conclusion Normoxic methane decreases the signs of oxidative and nitrosative stress, improves tissue microcirculation, and thus appears to modulate the ischemia-reperfusion–induced epithelial permeability changes. These findings suggest that the administration of exogenous methane may be a useful strategy for maintaining the integrity of the mucosa sustaining an oxido-reductive attack.

Background: Fractured neck of femur generally requires operative fixation and is a common cause of admission to hospital. The combination of femoral nerve block and spinal anesthesia is a common anesthetic technique used to facilitate the surgical procedure. The optimal disposition of local anesthetic (LA) relative the femoral nerve (FN) has not been defined. Our hypothesis was: that the deposition of LA relative to the FN influences the quality of analgesia for positioning of the patient for performance of spinal anesthesia. The primary outcome was verbal rating (VRS) pain scores 0–10 assessed immediately after positioning the patient to perform spinal anesthesia. Methods: With Institutional ethical approval and having obtained written informed consent from each, 52 patients were studied. The study was registered with ClinicalTrials.gov (NCT01527812). Patients were randomly allocated to undergo to one of three groups namely: intention to deposit lidocaine 2% (15 ml) i. above (Group A), ii. below (Group B), iii. circumferential (Group C) to the FN. A blinded observer assessed i. the sensory nerve block (cold) in the areas of the terminal branches of the FN and ii. VRS pain scores on passive movement from block completion at 5 minutes intervals for 30 minutes. Immediately after positioning the patient for spinal anesthesia, VRS pain scores were recorded. Results: Pain VRS scores during positioning were similar in the three groups [Above group/Below group/Circumferential group: 2(0–9)/0(0–10)/3(0–10), median(range), p:0.32]. The block was deemed to have failed in 20%, 47% and 12% in the Above group, Below group and Circumferential group respectively. The median number of needle passes was greater in the Circumferential group compared with the Above group (p:0.009). Patient satisfaction was greatest in the Circumferential group [mean satisfaction scores were 83.5(19.8)/88.1(20.5)/93.8(12.3), [mean(SD), p=0.04] in the Above, Below and Circumferential groups respectively. Conclusions: We conclude that there is no clinical advantage to attempting to deposit LA circumferential to the femoral nerve (relative to depositing LA either above or below the nerve), during femoral nerve block in this setting.