PLASMA PROTEIN LOSS DURING SURGERY: BENEFICIAL EFFECTS OF ALBUMIN SUBSTITUTION

Plasma protein loss during abdominal surgery is a known phenomenon, but its possible pathophysiological relevance has remained unknown. The present study evaluates the effects of albumin substitution on systemic and local hemodynamics and cellular interactions in the mesenteric microcirculation. Rats underwent median laparotomy and exteriorization of an ileal loop for intravital microscopy of the mesenteric microcirculation. Plasma protein concentrations, systemic and local hemodynamics were recorded during the follow up period, with or without albumin substitution. Depending on the time course of plasma protein loss in control experiments, 80% of the calculated protein loss was infused dur…

Description of the response of a new multi-parametric brain sensor to physiological and pathophysiological challenges in the cortex of juvenile pigs

AIM Monitoring of intracranial pressure (ICP), local cerebral blood flow (CBF) and oxygen is part of modern intensive critical care medicine. Preclinical evaluation of newly developed catheters that should monitor several parameters simultaneously is reported poorly in the literature. The goal of our study was (1) to evaluate a new multi-parametric sensor in brain tissue and (2) to establish a testing protocol using pathophysiological challenges that target measured parameters of the sensor and autoregulatory boundaries and could be used as preclinical standard protocol in future studies. MATERIAL AND METHODS We describe data from 12 new multi-parametric brain sensors (MPBS) that were impla…

Vitamin E supplements in standard rodent diets do not affect antioxidant intervention trials in apo-/-mice: failure of vitamin E to inhibit atherogenesis as assessed by histology in aortic arch and innominate/carotid arteries

A New Model of Skull Base Reconstruction following Expanded Endonasal or Transoral Approaches – Long-Term Results in Primates

<i>Objective:</i> The direct endonasal or transoral transclival approaches to the skull base permit effective minimally invasive surgery along the clivus region. Developing consistently effective techniques to prevent cerebrospinal fluid (CSF) leaks and their consequences (infections and healing processes with long and complicated recoveries) remains a major challenge. In this study, we tested over a long period a method of bone reconstruction newly developed by us, which makes use of a specially designed elastic silicone plug that can be employed for bone replacement after minimally invasive skull base surgery without risk of postoperative CSF leaks. After acute testing of plug…

Reperfusionsschock nach okklusion der A. mesenterica superior und akkumulation von leukozyten innerhalb der d�nndarmwand

Der Epithelschaden und die Leukozytenakkumulation in der Wand des Ileums nach Ischamie und Reperfusion wurden experimentell am Schwein untersucht. Die A. mesenterica superior wurde fur 1 h (Gruppe 2; n=9), 2 h (Gruppe 3; n=6) und 3 h (Gruppe 4; n=7) abgeklemmt und 2 h reperfundiert. Die histologische Beurteilung erfolgte an Hamatoxylin-Eosin- und NaphtolAS-D-Chlorazetatesterase-gefarbten Praparaten. Es entwickelte sich ein Reperfusionsschock in Abhangigkeit von der Ischamiedauer. Nach lstundiger Ischamie stabilisierte sich der Blutdruck mit erniedrigten Werten sowie einer Normalisierung des Serumlaktatspiegels und des intramuralen pHi des Dunndarms. Eine Verlangerung der Ischamie auf 2 h (G…

Surgical procedure affects physiological parameters in rat myocardial ischemia: need for mechanical ventilation.

Several surgical approaches are being used to induce myocardial ischemia in rats. The present study investigated two different operative procedures in spontaneously breathing and mechanically ventilated rats under sham conditions. A snare around the left coronary artery (LCA) was achieved without occlusion. Left lateral thoracotomy was performed in spontaneously breathing and mechanically ventilated rats (tidal volume 8 ml/kg) with a respiratory rate of 90 strokes/min at different levels of O2 supplementation (room air and 30, 40, and 90% O2). All animals were observed for 60 min after thoracotomy. Rats operated with exteriorization of the heart through left lateral thoracotomy while breat…

Low-Frequency Therapeutic Ultrasound with Varied Duty Cycle: Effects on the Ischemic Brain and the Inner Ear

Sonothrombolysis is a promising modality for acute stroke treatment. In vitro data suggest a duty cycle dependence of sonothrombolytic efficacy of low-frequency applications. The aim of our study was to examine its impact on safety issues in a rat model of middle cerebral artery occlusion. Rats were exposed to transcranial 60-kHz ultrasound with varied duty cycles. To determine effects on the inner ear, the acoustic threshold was determined in additional healthy animals (acoustic evoked potentials). A short duty cycle (20%) resulted in significant adverse effects (ischemic volume, hemorrhage, functional outcome), which was not observed in longer duty cycle (80%). Continuous-wave insonation …

Effects of a small acute subdural hematoma following traumatic brain injury on neuromonitoring, brain swelling and histology in pigs.

An acute subdural hematoma (ASDH) induces pathomechanisms which worsen outcome after traumatic brain injury, even after a small hemorrhage. Synergistic effects of a small ASDH on brain damage are poorly understood, and were studied here using neuromonitoring for 10 h in an injury model of controlled cortical impact (CCI) and ASDH. Pigs (n = 32) were assigned to 4 groups: sham, CCI (2.5 m/s), ASDH (2 ml) and CCI + ASDH. Intracranial pressure was significantly increased above sham levels by all injuries with no difference between groups. CCI and ASDH reduced ptiO<sub>2</sub> by a maximum of 36 ± 9 and 26 ± 11%, respectively. The combination caused a 31 ± 11% drop. ASDH alone and i…

Aufnahme und Phototoxizität von Hämatoporphyrinderivat in normalen und malignen Zellen

Die Photodynamische Therapie (PDT) stellt neben der Chemo-, Radio- und chirurgischen Therapie ein neues alternatives Therapiekonzept in der Behandlung von malignen Tumoren dar. Das Prinzip beruht auf der systemischen Applikation (i.v. Injektion) einer photosensibilisierenden Substanz, z.B. Hamatoporphyrinderivat (HPD), und der nachfolgenden Bestrahlung mit Licht geeigneter Wellenlange, z.B. Laserlicht (630 nm). Fur die tumorselektive Wirkung dieser Therapie werden die selektive Aufnahme bzw. vermehrte Speicherung der photoaktiven Substanzen im Tumorgewebe [1] sowie auch eine im Vergleich zum Normalgewebe erhohte Sensibuitat von Tumorgewebe gegenuber der PDT postuliert. Ungeklart ist, ob die…

Neuronal immunoreactivity for mannose-binding lectin after venous occlusion-induced focal cerebral ischemia in rats

Abstract A recent research reveals that complement activation exacerbates cerebral infarction. However, involvement of the lectin pathway, (the third complement activation pathway) in cerebral ischemia is not well studied. In this study, we investigated the appearance of mannose-binding lectin (MBL) in ischemic brain tissue. Male Wistar rats ( n  = 25) were divided into three groups: untreated control, sham, and vein occlusion (VO). Rats in the VO group had two adjacent photochemically occluded cortical veins. Regional cerebral blood flow (rCBF) was measured in the sham and VO groups. Rats were perfusion-fixed at 72 h in the sham group and at 3, 24, and 72 h after inducing ischemia in the V…

On the Number of Measurements Necessary to Assess Regional Cerebral Blood Flow by Local Laser Doppler Recordings: A Simulation Study with Data from 45 Rabbits

Laser Doppler fluxmetry may improve the monitoring of cortical blood flow in neurosurgical patients. So far, however, the variability of laser Doppler readings found in the cerebral cortex has prevented a consequent usage of the technique in clinical practice. The current report analyzes the regional variability of laser Doppler readings from experimental animals. Typical frequency histograms of observed flow values display non-Gaussian distributions. A simulation technique is used to assess the number of measuring sites required for valid estimates of regional cortical flow. From a total of 990 local flow measurements from 45 rabbits random samples of sizes between 5 and 100 were repeatedl…

Alterations of regional cerebral blood flow and oxygen saturation in a rat sinus-vein thrombosis model.

Background and Purpose The pathophysiology of sinus-vein thrombosis (SVT) in patients and experimental animals is still poorly understood. This study was designed to examine and further elucidate the pathophysiological sequence of events, especially the relationship between local and regional blood flow and hemoglobin oxygen saturation (HbSO 2 ) detected at identical locations. The use of both parameters as outcome indicators should be compared. Methods SVT was induced by ligation of the superior sagittal sinus (SSS) and slow injection of kaolin-cephalin suspension into the SSS in rats. Regional cerebral blood flow (rCBF) was assessed by laser-Doppler flowmetry together with regional HbSO …

Effects of hypertonic/hyperoncotic treatment after rat cortical vein occlusion*

Objective To examine the effects of hypertonic/hyperoncotic treatment on physiologic variables and regional cerebral blood flow and to test its neuroprotective efficiency in a model of permanent venous ischemia. Design Randomized prospective study. Setting University research institute. Subjects Adult male Wistar rats, weighing 359 +/- 54 g (n = 38). Interventions Rats were subjected to photochemical occlusion of two adjacent cortical veins. A randomized infusion with vehicle (0.9% NaCl), 10% hydroxyethyl starch 200,000 (HES), or 7.5% saline plus 10% hydroxyethyl starch 200,000 (HHES) was started 30 mins after two-vein occlusion. Effects on physiologic variables and regional cerebral blood …

Moderate controlled cortical contusion in pigs: effects on multi-parametric neuromonitoring and clinical relevance.

Over the last decade, routine neuromonitoring of ICP and CPP has been extended with new on-line techniques such as microdialysis, tissue oxygen (ptiO(2)), acid-base balance (ptiCO(2), pH) and CBF measurements, which so far have not lead to clear-cut therapy approaches in the neurointensive care unit. This is partially due to the complex pathophysiology following a wide-range of brain injuries, and the lack of suitable animal models allowing simultaneous, clinically relevant neuromonitoring under controlled conditions. Therefore, a controlled cortical impact (CCI) model in large animals (pig) has been developed. After placement of microdialysis, ptiO(2), temperature and ICP catheters, an uni…

On the number of measurement sites required to assess regional cerebral blood flow by laser-Doppler scanning during cerebral ischemia and reperfusion.

The aim of this study was to determine whether the number of measurement sites affected the precision of regional cerebral blood flow (CBF) assessment by Laser-Doppler (LD). A simulation study was applied based on data obtained by scanning the cortex in 25 rats during baseline conditions, 15 min global cerebral ischemia and reperfusion. Random samples were repeatedly collected from 1 to 100 locations and deviations from the median of the entire CBF data pool (800 locations) were determined. Single location CBF measurements missed the true median by 24.8+/-2.2 LD-units (baseline conditions, n=100 simulations, mean+/-SEM), 2.7+/-0.6 LD-units (ischemia), and 31.9+/-2.4 LD-units (30th min reper…

Sub-cellular tumor identification and markerless differentiation in the rat brain in vivo by multiphoton microscopy

Objective/Background Aim of the current study was to localize and differentiate between tumor (glioma) and healthy tissue in rat brains on a cellular level. Near-infrared multiphoton microscopy takes advantage of the simultaneous absorption of two or more photons to analyze various materials such as cell and tissue components via the observation of endogenous fluorophores such as NAD(P)H, FAD, porphyrins, melanin, elastin, and collagen, with a very high resolution, without inducing the problems of photo-bleaching on out-of-focus areas. Methods In vitro and in vivo studies on healthy rat brains as well as C6 glioma cell line allografts have been performed. Near-infrared laser pulses (λ = 690…

Pharmacological preconditioning in global cerebral ischemia

Single dose 3-nitropropionic acid (3-NPA) 24 hr before global ischemia improves neuronal survival in both, neocortex and hippocampus (‘chemical preconditioning’). Neuronal survival after transient global ischemia requires new protein synthesis during recovery, especially of those with anti-apoptotic function. Bcl-2-protein is expressed in neurons that survive cerebral ischemia and may parallel the time course of tolerance after ischemic preconditioning. With this study we examined whether differences in bcl-2-protein expression compared to baseline may be involved in the induction of ischemic tolerance using 3-NPA.

Using multidimensional scaling in model choice for congenital oesophageal atresia: similarity analysis of human autopsy organ weights with those from a comparative assessment of Aachen Minipig and Pietrain piglets.

Swine models had been popular in paediatric oesophageal surgery in the past. Although being largely replaced by rodent models, swine experienced a revival with the establishment of minipig models. However, none of them has ever been investigated for similarity to humans. We conducted a pilot study to determine whether three-week old Pietrain piglets and three-month old Aachen Minipigs are suitable for experimental paediatric oesophageal atresia surgery. We tested the operation's feasibility, performed a necropsy, weighed organs, measured organ length and calculated relative weights and lengths, and measured laboratory parameters. We used multidimensional scaling to assess the similarity of…

Modelling of the Ischemic Penumbra

What happens to the ischemic penumbra – defined as a territory of critically reduced blood flow in the close neighborhood of an ischemic core – determines outcome after stroke. Currently the pathophysiology of the penumbra is studied predominantly in rat models with occlusion of the middle cerebral artery. Here we propose two other rat models with distinct advantages. One produces a large territory of critical flow reduction in the cortex of one hemisphere without presence of an infarct core: this model is suited to study mediator mechanisms that may transform the penumbra into necrotic tissue. It is produced by occluding one carotid artery and in addition reducing arterial pressure to 50mm…

Selective pelvic autonomic nerve stimulation with simultaneous intraoperative monitoring of internal anal sphincter and bladder innervation.

<i>Background:</i> Pelvic autonomic nerve preservation avoids postoperative functional disturbances. The aim of this feasibility study was to develop a neuromonitoring system with simultaneous intraoperative verification of internal anal sphincter (IAS) activity and intravesical pressure. <i>Methods:</i> 14 pigs underwent low anterior rectal resection. During intermittent bipolar electric stimulation of the inferior hypogastric plexus (IHP) and the pelvic splanchnic nerves (PSN), electromyographic signals of the IAS and manometry of the urinary bladder were observed simultaneously. <i>Results:</i> Stimulation of IHP and PSN as well as simultaneous intraop…

An experimental study on long term outcomes after magnetic esophageal compression anastomosis in piglets

Abstract Background/Purpose Previous studies have shown that a patent, watertight esophageal anastomosis can be accomplished safely using specially-shaped magnets in piglets. However, it is unclear whether such a magnetic esophageal compression anastomosis (MECA) remains patent in the long-term. The purpose of this study was to evaluate the long-term outcome of MECA in an experimental pig model over an observation period of 2 months. Methods Ten piglets underwent creation of an MECA with custom-made 8 mm magnets and a U-shaped esophageal bypass loop to allow peroral nutrition at eight weeks of life. Two weeks later, the bypass loop was closed surgically, requiring the pigs to swallow via th…

An in vitro model to study cellular photosensitizer uptake and photodynamic dose-response relationships of tumor cells

Cellular fluorescence intensity (CFI) after incubation with varying concentrations of the photosensitizer Photofrin and the photodynamically induced dose-response relationships of hamster melanoma cells (A-MEL-3) were studied in a recently developed in vitro model. After administration of Photofrin to the extracellular serum-free medium, CFI was evaluated by flow cytometry together with constantly fluorescing latex particles used as a reference. After 5 min, 50% of maximal CFI was found, and after 60 min CFI was maximal. No further increase was obtained during the exposure to Photofrin over the incubation period of 4 h. During this plateau phase, CFI was significantly related to the concent…

Comparison of the quantitative first pass myocardial perfusion MRI with and without prospective slice tracking: Comparison between breath-hold and free-breathing condition

Physiologic motion of the heart is one of the major problems of myocardial blood flow quantification using first pass perfusion-MRI method. To overcome these problems, a perfusion pulse sequence with prospective slice tracking was developed. Cardiac motion was monitored by a navigator directly positioned at heart's basis to overcome no additional underlying model calculations connecting diaphragm and cardiac motion. Additional prescans were used before the perfusion measurement to detect slice displacements caused by remaining cardiac motion between navigator and the perfusion slice readout. The pulse sequence and subsequent quantification of myocardial blood flow was tested in healthy pigs…

In vivo imaging of dopamine receptors in a model of temporal lobe epilepsy

Alterations in dopamine neurotransmission in animal models of epilepsies have been frequently demonstrated using invasive neuroscience or ex vivo techniques. We aimed to test whether corresponding alterations could be detected by noninvasive in vivo brain imaging with positron emission tomography (PET) in the chronic phase of the rat pilocarpine model of temporal lobe epilepsy.Six pilocarpine-treated Wistar rats exhibiting spontaneous recurrent seizures and nine control rats were studied with PET using [(18)F]-fallypride, a high-affinity dopamine D(2/3) receptor ligand. Parametric images of [(18)F]-fallypride specific binding were calculated using a reference tissue method, and the two grou…

Intrahematomal Ultrasound Enhances RtPA-Fibrinolysis in a Porcine Model of Intracerebral Hemorrhage

Catheter-based ultrasound-thrombolysis has been successfully used in a small clinical trial in order to enhance recombinant tissue plasminogen activator (rtPA)-fibrinolysis, for the treatment of spontaneous intracerebral hemorrhages (ICHs). The aim of this study was to investigate the ultra-early effects of ultrasound on hematoma and the surrounding brain tissue in a porcine ICH-model. To achieve this, 21 pigs with a right frontal ICH were randomly assigned to four groups: (1) drainage (n = 3), (2) drainage + rtPA (n = 6), (3) drainage + ultrasound (n = 6), and (4) drainage + ultrasound + rtPA (n = 6). The hematoma volume assessment was performed using cranial MRI before and after the treat…

Blood constituents trigger brain swelling, tissue death, and reduction of glucose metabolism early after acute subdural hematoma in rats

Outcome from acute subdural hematoma is often worse than would be expected from the pure increase of intracranial volume by bleeding. The aim was to test whether volume-independent pathomechanisms aggravate damage by comparing the effects of blood infusion with those of an inert fluid, paraffin oil, on intracranial pressure (ICP), cerebral perfusion pressure (CPP), local cerebral blood flow (CBF), edema formation, glucose metabolism ([18F]-deoxyglucose, MicroPET), and histological outcome. Rats were injured by subdural infusion of 300 μL venous blood or paraffin. ICP, CPP, and CBF changes, assessed during the first 30 mins after injury, were not different between the injury groups at most …

Low-frequency ultrasound induces nonenzymatic thrombolysis in vitro.

To evaluate whether ultrasound, applied over a distance of several centimeters and in the absence of thrombolytic agents, may have a thrombolytic effect on blood clots.Low-frequency (20 kHz) continuous wave ultrasound at different intensity levels (0.15-1.2 W/cm2) and exposure times (5, 10, and 20 minutes) was assessed for its potential to induce thrombolysis of fresh human blood clots. The ultrasound effect was also studied in combination with recombinant tissue-type plasminogen activator-mediated thrombolysis. Experiments were carried out in a flow model in degassed sodium phosphate buffer at 37 degrees C at a distance of 3 cm from the ultrasonic probe to the blood clots. Regardless of ul…

An early bolus of hypertonic saline hydroxyethyl starch improves long-term outcome after global cerebral ischemia.

Objective: The beneficial effect of hypertonic saline solutions in the emergency treatment of shock and traumatic brain injury is well described. The present study determines effects of a single bolus of hypertonic saline on long-term survival, neurologic function, and neuronal survival 10 days after global cerebral ischemia. In addition, we evaluated the therapeutic window for hypertonic saline treatment (early vs. delayed application). Design: Laboratory experiment. Setting: University laboratory. Subjects: Male Wistar rats weighing 240‐330 g. Interventions: Rats were submitted to temporal global cerebral ischemia using temporary bilateral carotid occlusion combined with hypobaric hypoten…

Electrical stimulation with motility analysis of five parts of the gastrointestinal tract ; First evaluation of an experimental protocol in a porcine model

Abstract Gastrointestinal (GI) motility disorders are frequent and clinically significant conditions with impairment of patient’s quality of life. Examples range from upper GI symptoms such as dysphagia and gastroparesis to lower GI manifestations, namely chronic-intestinal pseudo obstruction, diarrhea and constipation. Furthermore, postoperative motility disorders are common. Currently, available pharmacological or dietetic treatment options are limited. Since GI motility is based on myoelectric activity, electrical stimulation (ES) is a promising alternative. Numerous studies have demonstrated suitable pacing strategies and parameters in different GI segments. However, results of multiloc…

Mapping brain activity with flexible graphene micro-transistors

arXiv:1611.05693v1.-- et al.

New method of bone reconstruction designed for skull base surgery

The direct endonasal or transoral transclival approaches to the skull base permit effective, minimally invasive surgery along the clivus. Developing long-term, effective techniques to prevent cerebrospinal fluid (CSF) leaks and their consequences (infection and delayed healing) remains a major challenge. In this study we describe a method of bone reconstruction newly developed by us, which uses a custom designed silicone plug for bone replacement after minimally invasive skull base surgery with a low incidence of postoperative CSF leaks. German Landrace pigs were used to test the efficiency of the new technique. Twelve craniotomies were performed in six pigs using a subtemporal approach and…

A Continuous Suture Anastomosis Outperforms a Simple Interrupted Suture Anastomosis in Esophageal Elongation.

Abstract Introduction Long-gap esophageal atresia represents a distinct entity among the esophageal atresia spectrum. In many patients, achieving a reasonable anastomosis depends on some millimeters of tissue. We aimed to determine what effect the suturing technique would have on esophageal ex vivo elongation as it may determine the strength of a primary anastomosis. Materials and Methods In an analysis of porcine esophagi from animals for slaughter (100–120 days old with a weight of 100–120 kg), we determined esophageal length gain of simple continuous and simple interrupted suture anastomoses subjected to linear traction until linear breaking strength was reached. Statistical power of 80%…

Microcirculation after cerebral venous occlusions as assessed by laser Doppler scanning.

✓ Research on cerebral venous circulation disturbances (CVCDs) has been limited partly by the paucity of animal models that produce consistent venous infarction. Occlusion of two adjacent cortical veins in rats by means of a photochemical thrombotic technique provides a minimally invasive, clinically relevant, and reproducible model suited to study the pathophysiology of CVCDs. In this study, the effects of venous occlusion on regional cortical blood flow and the brain damage that ensues were evaluated. Cortical vein occlusion was induced by photoactivation of rose bengal via 100-µm fiberoptic illumination. The cerebral venous flow pattern was examined using fluorescence angiography until 9…

Kinetics of Photofrin II in perifocal brain edema.

Photodynamic therapy is under intense investigation as a possible adjuvant for the treatment of malignant tumors of the central nervous system. It relies on the fact that photosensitizers are selectively taken up or retained by malignant tissue. However, most brain tumors are accompanied by substantial vasogenic edema as a consequence of blood-brain barrier disruption within the tumor, leading to extravasation and propagation of plasma constituents into the surrounding brain tissue. Systemically administered photosensitizers may enter healthy tissue together with the edema fluid, possibly leading to sensitization of tissues outside the tumor. To test this hypothesis, vasogenic edema was ind…

Temporal profile of gene induction after venous ischemia accompanied by spreading depression as compared to spreading depression alone

Abstract The occlusion of two adjacent cortical veins is followed by a widespread reduction of rCBF and the occurrence of small infarcts, which become larger if spreading depression (SD) occurs. The model appears to be useful for studying penumbra pathophysiology. Here, the time course of gene expression in the penumbra was compared to those induced by the SD alone. In an experiment involving rats, an occlusion of two cortical veins was induced by i.v. rose bengal and fiberoptic illumination. Ten SDs were induced at 7-min intervals. Changes of the gene expression after 2, 8, 24 and 72 h were analysed for 13 genes by RT-PCR. The housekeeping gene glyceraldehyde-3-phosphate dehydrogenase (GAP…

Tightness of duraplasty in rabbits: a comparative study.

OBJECTIVE: The purpose of this study was to test, in rabbits, the tightness of seven dural substitution materials commonly used in neurosurgery, i.e., Lyodura (B. Braun Melsungen AG, Melsungen, Germany), Tutoplast dura (Tutogen Medical, Inc., Parsippany, NJ), Tutoplast fascia lata (Tutogen Medical, Inc.), autologous periosteum, Neuropatch (B. Braun Melsungen AG), Dacron (E.I. du Pont de Nemours and Co., Wilmington, DE), and Ethisorb (Ethicon, Inc., Somerville, NJ). METHODS: Duraplasties were performed with sutures alone or were additionally fixed with fibrin glue. Leakage pressures were assessed by infusion of artificial cerebrospinal fluid, containing sodium fluorescein, into the cisterna …

MitoKATP-channel opener protects against neuronal death in rat venous ischemia.

OBJECTIVE: Mitochondrial adenosine triphosphate-dependent potassium (mitoK ATP ) channels are present in the brain, and several reports have shown their neuroprotective, preconditioning effect against an ischemic insult. The role of mitoK ATP channels in the penumbra area has not been studied thoroughly. In a model of venous ischemia, widespread penumbra-like low flow areas are created, which are susceptible to cortical spreading depression. Thus, we studied effects of mitoK ATP channels on infarct size in this model. METHODS: Male Wistar rats were subjected to two-vein occlusion by photochemical thrombosis of two adjacent cortical veins combined with KCI-induced cortical spreading depressi…

Capillary flow and diameter changes during reperfusion after global cerebral ischemia studied by intravital video microscopy.

The reaction of cerebral capillaries to ischemia is unclear. Based on Hossmann's observation of postischemic “delayed hypoperfusion,” we hypothesized that capillary flow is decreased during reperfusion because of increased precapillary flow resistance. To test this hypothesis, we measured cerebral capillary erythrocyte velocity and diameter changes by intravital microscopy in gerbils. A cranial window was prepared over the frontoparietal cortex in 26 gerbils anesthetized with halothane. The animals underwent either a sham operation or fifteen minutes of bilateral carotid artery occlusion causing global cerebral ischemia. Capillary flow velocities were measured by frame-to-frame tracking of…

Temporal and Spatial Evolution of Cortical Blood Flow in the Penumbra of Venous Infarcts

A cortical venous infarction model has been evaluated regarding the temporal and spatial evolution of regional flow reduction. Two cortical veins were occluded photochemically with rose bengal and fiberoptic illumination. Seven rats served to demonstrate effects on regional cortical blood flow using laser Doppler scanning. After two-vein occlusion, there was a widespread reduction of flow that gradually deteriorated after vein occlusion when regional cortical flow in a 3.5 x 7.0 mm window: after 15 min it had decreased to 57.8% ± 8.0%, and after 75 min it was 34.3% ± 5.4%. Infarct volumes as determined in 10 rats 5 days after two-vein occlusion had an average size of 3.6 ± 0.7 mm3. The data…

Concept and setup for intraoperative imaging of tumorous tissue via Attenuated Total Reflection spectrosocopy with Quantum Cascade Lasers

A major challenge in tumor surgery is the differentiation between normal and malignant tissue. Since an incompletely resected tumor easily leads to recidivism, the gold standard is to remove malignant tissue with a sufficient safety margin and send it to pathology for examination with patho-histological techniques (rapid section diagnosis). This approach, however, exhibits several disadvantages: The removal of additional tissue (safety margin) means additional stress to the patient; the correct interpretation of proper tumor excision relies on the pathologist’s experience and the waiting time between resection and pathological result can be more than 45 minutes. This last aspect implies unn…

Cerebral blood flow autoregulation during hypobaric hypotension assessed by laser Doppler scanning.

Hypobaric hypotension was used to reduce systemic blood pressure in rats below the lower threshold of CBF autoregulation to evaluate a new laser Doppler (LD) “scanning” technique. Spontaneously breathing male Wistar Kyoto rats (n = 8) were anesthetized with chloral hydrate and the head fixed in a stereotaxic head holder. A cranial window with intact dura mater was introduced to assess local CBF (lCBF) by LD. One stationary probe served to detect rapid flow changes, whereas the second probe was used to sample lCBF recordings from many cortical locations by means of a stepping motor-controlled micromanipulator to obtain lCBF frequency histograms. Advantages are an improved spatial resolution…

Neuroprotective effect of ceftriaxone on the penumbra in a rat venous ischemia model.

Glutamate transporter-1 (GLT-1) maintains low concentrations of extracellular glutamate by removing glutamate from the extracellular space. It is controversial, however, whether upregulation of GLT-1 is neuroprotective under all ischemic/hypoxic conditions. Recently, a neuroprotective effect of preconditioning with a β-lactam antibiotic ceftriaxone (CTX) that increases expression of GLT-1 has been reported in animal models of focal ischemia. On the other hand, it is said that CTX does not play a neuroprotective role in an in vitro study. Thus, we examined the effect of CTX on ischemic injury in a rat model of two-vein occlusion (2VO). This model mimics venous ischemia during, e.g. tumor sur…

Summary Background. Photodynamic therapy (PDT) has been under dis- cussion as additional treatment option for malignant gliomas. How- ever, damage not only to tumour tissue but also to normal brain has been demonstrated. The mechanisms of this unwanted side eect have not yet been clearly identified. Spreading of photosensitiser with oedema after disruption of the blood-brain-barrier and poten- tial sensitisation of normal tissue has been foundpreviously. The present study investigates the time- and dose-dependency of normal tissue damage to photodynamic therapy using Photofrin II9 after disruption of the blood-brain-barrier. Methods. Male wistar rats anaesthetisedwith chloral hydrate were s…

Caspase-dependent cell death involved in brain damage after acute subdural hematoma in rats

Abstract Traumatic brain injury is associated with acute subdural hematoma (ASDH) that worsens outcome. Although early removal of blood can reduce mortality, patients still die or remain disabled after surgery and additional treatments are needed. The blood mass and extravasated blood induce pathomechanisms such as high intracranial pressure (ICP), ischemia, apoptosis and inflammation which lead to acute as well as delayed cell death. Only little is known about the basis of delayed cell death in this type of injury. Thus, the purpose of the study was to investigate to which extent caspase-dependent intracellular processes are involved in the lesion development after ASDH in rats. A volume o…

C1-Esterase-Inhibitor Treatment at Early Reperfusion of Hemorrhagic Shock Reduces Mesentery Leukocyte Adhesion and Rolling

Objective: Complement activation probably plays a pathogenic role in multiple organ failure in shock. This study evaluates the effects of C1-esterase-inhibitor treatment on leukocyte-endothelial interaction in the mesenteric microcirculation in hemorrhagic shock. Methods: Rats underwent median laparotomy and exteriorization of an ileal loop for intravital microscopy of the mesenteric microcirculation. Volume controlled hemorrhagic shock was provoked by arterial blood withdrawal (2.5 mL/ 100 g body wt. for 60 minutes) followed by a 4-hour reperfusion period. C1-INH (100 IU/kg body wt. i.v.) or 0.9% NaCl i.v. were administered as a bolus at the beginning of reperfusion. Reperfusion time mimic…

C1-esterase inhibitor reduces infarct volume after cortical vein occlusion.

In order to clarify the role of complement as a mediator of cerebral infarct growth, we inhibited the classical complement activation pathway in a photochemical cortical vein occlusion model. Immediately after occlusion, rats were infused with either 0.9% saline (vehicle), or C1-esterase inhibitor (C1-INH) over 30 min. Regional cerebral blood flow (rCBF) decreased after occlusion, and was about 50% of baseline after 2 h. No difference was noted between experimental groups. Mean arterial blood pressure (MABP) and arterial blood gases were likewise unaffected by the treatment. However, administration of C1-INH had significantly reduced infarct volume by 72%, as evaluated after 5 days survival…

Early-onset tolerance in rat global cerebral ischemia induced by a mitochondrial inhibitor

It was studied whether a subtoxic dose of the mitochondrial neurotoxin, 3-nitropropionic acid (3-NPA), can initiate early-onset tolerance induction for subsequent ischemic injury. Wistar rats were pretreated for 3 h by intraperitoneal 3-NPA (20 mg/kg body weight; n=13) or solvent (n=12). Fifteen minutes global cerebral ischemia was induced by bilateral carotid artery occlusion and hypobaric hypotension. rCBF and tissue hemoglobin oxygen saturation were measured by laser Doppler scanning and a microspectrophotometric method. Ischemic insult and brain temperature were identical in both groups. Body weight and neurological scores recovered in the pretreated group but further deteriorated in th…

Acute Subdural Hematoma in Pigs: Role of Volume on Multiparametric Neuromonitoring and Histology

Traumatic brain injury (TBI) is often complicated by acute subdural hemorrhage (ASDH) with a high mortality rate. The pathophysiological mechanisms behind such an injury type and the contribution of blood to the extent of an injury remain poorly understood. Therefore, the goals of this study were to establish a porcine ASDH model in order to investigate pathomechanisms of ASDH and to compare effects induced by blood or sheer volume. Thus, we infused 2, 5, and 9 mL of blood (up to 15% of intracranial volume), and we compared a 5-mL blood and paraffin oil volume to separate out effects of extravasated blood on brain tissue. An extended neuromonitoring was applied that lasted up to 12 h after …

Erythropoietin neuroprotection is enhanced by direct cortical application following subdural blood evacuation in a rat model of acute subdural hematoma.

Recombinant human erythropoietin (EPO) has been successfully tested as neuroprotectant in brain injury models. The first large clinical trial with stroke patients, however, revealed negative results. Reasons are manifold and may include side-effects such as thrombotic complications or interactions with other medication, EPO concentration, penetration of the blood-brain-barrier and/or route of application. The latter is restricted to systemic application. Here we hypothesize that EPO is neuroprotective in a rat model of acute subdural hemorrhage (ASDH) and that direct cortical application is a feasible route of application in this injury type. The subdural hematoma was surgically evacuated a…

Temporal Profile of Gene Induction After Venous Ischemia and Effects of Spreading Depression

Occlusion of two adjacent cortical veins is followed by a widespread reduction of rCBF and the occurrence of small infarcts, which become larger if spreading depression (SD) occurs. The infarct matures over time with TUNEL-positive cells seen in the penumbra up to 4 days after vein occlusion. Caspase inhibition with zVAD.fmk reduces infarct size. Here, the time course of gene expression in the penumbra is compared to that induced by SD alone.

Bodyweight, not age, determines oesophageal length and breaking strength in rats

Delayed primary repair is still the method of choice in the management of long-gap oesophageal atresia in many centres, but the timing of anastomoses varies. Some assume the infant's bodyweight to be an important factor, whereas others prefer age. We therefore aimed to clarify whether age or bodyweight determined oesophageal length in a rodent model.We explanted the oesophagi of 20 Sprague-Dawley rats, aged 15 to 444 days (n = two per time point), measured bodyweight, oesophageal length, weight, and linear breaking strength to measure tissue resilience. Univariate and multivariate regression analyses were conducted to determine the influence of age and bodyweight on oesophageal length and l…

Spontaneous Cortical Spreading Depression and Intracranial Pressure Following Acute Subdural Hematoma in a Rat

Acute subdural hemorrhage (ASDH) is a frequent and devastating consequence of traumatic brain injury. Tissue damage develops rapidly and makes treatment even more difficult. Management of increased intracranial pressure (ICP) due to extravasated blood volume and brain swelling is often insufficient to control all adverse effects of ASDH. In addition to sheer volume, spontaneously triggered cortical spreading depression (CSD) that leads to cell death following ischemia or trauma may contribute to injury development after ASDH. Therefore, we explored the occurrence of CSD by tissue impedance (IMP) measurement in a rat model subjected to ASDH. IMP and intraventricular and mean arterial pressur…

Occlusion of the pig superior sagittal sinus, bridging and cortical veins: multistep evolution of sinus-vein thrombosis

✓ Cerebral sinus-vein thrombosis may lead to severe hemodynamic changes, elevated intracranial pressure (ICP), and brain edema. It is supposed that progression of the thrombus from the sinus into bridging and cortical veins plays a key role in the development of these pathophysiological changes, but this hypothesis lacks experimental proof. The aim of this study, using a novel animal model of sinus-vein thrombosis, was to evaluate the effects of a standardized occlusion of the superior sagittal sinus and its bridging and cortical veins on hemodynamic alterations, on brain water content, and on ICP in domestic pigs. In 10 animals, the middle third of the superior sagittal sinus was occluded …

Five-fold Gastrointestinal Electrical Stimulation With Electromyography-based Activity Analysis: Towards Multilocular Theranostic Intestinal Implants

Background/aims Motility disorders are common and may affect the entire gastrointestinal (GI) tract but current treatment is limited. Multilocular sensing of GI electrical activity and variable electrical stimulation (ES) is a promising option. The aim of our study is to investigate the effects of adjustable ES on poststimulatory spike activities in 5 GI segments. Methods Six acute porcine experiments were performed with direct ES by 4 ES parameter sets (30 seconds, 25 mA, 500 microseconds or 1000 microseconds, 30 Hz or 130 Hz) applied through subserosal electrodes in the stomach, duodenum, ileum, jejunum, and colon. Multi-channel electromyography of baseline and post-stimulatory GI electri…

Transcutaneous sacral nerve stimulation for intraoperative verification of internal anal sphincter innervation.

Background: The current standard for pelvic intraoperative neuromonitoring (pIONM) is based on intermittent direct nerve stimulation. This study investigated the potential use of transcutaneous sacral nerve stimulation for non‐invasive verification of pelvic autonomic nerves. Methods: A consecutive series of six pigs underwent low anterior rectal resection. For transcutaneous sacral nerve stimulation, an array of ten electrodes (cathodes) was placed over the sacral foramina (S2 to S4). Anodes were applied on the back, right and left thigh, lower abdomen, and intra‐anally. Stimulation using the novel method and current standard were performed at different phases of the experiment…

Postischemic application of lipid peroxidation inhibitor U-101033E reduces neuronal damage after global cerebral ischemia in rats.

Background and Purpose —The lipid peroxidation inhibitor U-101033E was examined for effects on cerebral blood flow (CBF), cortical tissue hemoglobin oxygen saturation (HbS o 2 ), and neuronal damage. Methods —Fifteen minutes of global cerebral ischemia was induced by two-vessel occlusion and hypobaric hypotension. Wistar rats (n=25) were randomized to receive vehicle (n=9) or 40 mg/kg U-101033E (n=9) intraperitoneally during 2 hours of reperfusion. A sham group (n=7) had neither ischemia nor therapy. Histology was evaluated 7 days after ischemia. Results —During late hyperperfusion (at 17 minutes), vehicle-treated animals had a higher ( P =0.044) cortical tissue HbS o 2 (72.0±1.4%) than di…

Intracoronary application of C1 esterase inhibitor improves cardiac function and reduces myocardial necrosis in an experimental model of ischemia and reperfusion.

Background Myocardial injury from ischemia can be aggravated by reperfusion of the jeopardized area. The precise underlying mechanisms have not been clearly defined, but proinflammatory events, including complement activation, leukocyte adhesion, and infiltration and release of diverse mediators, probably play important roles. The present study addresses the possibility of reducing reperfusion damage by the application of C1 esterase inhibitor (C1-INH). Methods and Results Cardioprotection by C1-INH 20 IU/kg IC was examined in a pig model with 60 minutes of coronary occlusion, followed by 120 minutes of reperfusion. C1-INH was administered during the first 5 minutes of coronary reperfusion…

Comparing Aachen Minipigs and Pietrain Piglets as Models of Experimental Pediatric Urology to Human Reference Data

<b><i>Introduction:</i></b> Swine had special roles in the development of minimally invasive procedures to treat vesicoureteral reflux, and minipigs have been gaining ground in recent years in experimental pediatric urology as they combine small size with less vulnerable adult physiology, but their suitability as a model has never been assessed. We therefore compared a landrace piglet with a juvenile minipig to elucidate comparability. <b><i>Methods:</i></b> We evaluated five 3-week old Pietrain piglets and five 3-month old Aachen Minipigs as representatives of landrace and minipig models based on their expected bodyweight being similar to a n…

The neuroprotective effect of lactate is not due to improved glutamate uptake after controlled cortical impact in rats.

For many years lactate was considered to be a waste product of glycolysis. Data are accumulating that suggest that lactate is an important energy substrate for neurons during activation. In severe traumatic brain injury (TBI) glutamate release and ischemic cerebral blood flow (CBF) are major factors for a mismatch between energy demand and supply and for neuronal cell death. Although ATP and behavior could be improved by lactate treatment after TBI, no histological correlate nor any linkage to better astrocytic glutamate uptake or CBF as possible mechanisms have been described. We subjected male rats to a controlled cortical impact (CCI; 5 m/sec, 2.5 mm). To study the effects of lactate tre…

“Small Volume Resuscitation” as Treatment of Cerebral Blood Flow Disturbances and Increased ICP in Trauma and Ischemia

“Small volume resuscitation” (SVR) is a promising concept for the treatment of shock and trauma patients. SVR utilizes the fast infusion of a small volume of hypertonic saline to mobilize intraendothelial and parenchymal water to expand and restitute intravascular volume. Therefore it seems warranted to also consider SVR for the treatment of disturbances of the cerebral circulation and of increased intracranial pressure (ICP). The current study uses a rabbit model of global cerebral ischemia combined with mild hemorrhage to test SVR. Somatosensory evoked potentials (SEPs) serve as a short-term outcome parameter. The data demonstrate a beneficial effect on ICP, a better reperfusion, and an i…

Resection of C6 gliomas in rats with the aid of the waterjet technique

While clinically the safety and efficacy of waterjet resection of brain tumors have been shown, evidence that waterjet dissection improves tumor resection radicality in comparison with conventional techniques is still missing. In the present study, resection radicality and tumor-free long-term survival of both techniques were evaluated in a C6-glioma model.Fifty-thousand C6-glioma cells were stereotactically transplanted in the left frontal lobe of 100 male Sprague-Dawley rats. After MRI-scanning for evaluation of tumor extension, microsurgical tumor resection was performed with conventional techniques (n=50) or with the waterjet dissector at pressures of 6bar (n=50). Twenty-five animals of…

Monitoring of cortical blood flow: Clinical relevance of experimental laser Doppler studies

Continuous monitoring of cortical blood flow provides real time information of CBF-changes during neurosurgical operations and on the neurointensive care unit. Laser Doppler flowmetry is a continuous, noninvasive technique suitable for measurement of the cortical microcirculation. In a number of experimental studies we have analyzed the characteristics of this method for cortical blood flow monitoring. The high spatial resolution of laser Doppler flowmetry and the heterogeneity of the cortical microvascular network results in a scatter of flow values over a wide range depending on the site of measurement. Data collection from different spots and calculation of frequency histogram may serve …

Crucial role for Nox2 and sleep deprivation in aircraft noise-induced vascular and cerebral oxidative stress, inflammation, and gene regulation

Abstract Aims Aircraft noise causes endothelial dysfunction, oxidative stress, and inflammation. Transportation noise increases the incidence of coronary artery disease, hypertension, and stroke. The underlying mechanisms are not well understood. Herein, we investigated effects of phagocyte-type NADPH oxidase (Nox2) knockout and different noise protocols (around-the-clock, sleep/awake phase noise) on vascular and cerebral complications in mice. Methods and results C57BL/6j and Nox2 −/− (gp91phox −/−) mice were exposed to aircraft noise (maximum sound level of 85 dB(A), average sound pressure level of 72 dB(A)) around-the-clock or during sleep/awake phases for 1, 2, and 4 days. Adverse effec…

Cluster analysis of mrna expression levels identifies multiple sequential patterns following focal cerebral ischemia

AIM The purpose of this study is to detect gene expression patterns following focal cerebral ischemia. MATERIAL AND METHODS 25 male Wistar rats were divided into control (n = 8) and ischemic (n = 17) groups. In the ischemic group, slowly progressing focal ischemia was simulated by two-vein occlusion with spreading depression (SD) a cortical microinjection of KCl induced. Ischemic tissue was removed at 2, 8, 24, or 72 h postischemia. Using semiquantitative reverse transcription polymerase chain reaction, we investigated mRNA expression levels of 13 representative genes related to cerebral ischemia. Cluster analysis of the gene expression levels was done. RESULTS In the ischemic group, the ex…

Online signal processing of internal anal sphincter activity during pelvic autonomic nerve stimulation: a new method to improve the reliability of intra-operative neuromonitoring signals

Aim  Intra-operative neuromonitoring is increasingly applied in several surgical disciplines and has been introduced to facilitate pelvic autonomic nerve preservation. Nevertheless, it has been considered a questionable tool for the minimization of risk, as the results are variable and might be misleading. The aim of the present experimental study was to develop an intra-operative neuromonitoring system with improved reliability for monitoring pelvic autonomic nerve function. Method  Fifteen pigs underwent low anterior rectal resection with pelvic autonomic nerve preservation. Intra-operative neuromonitoring was performed under autonomic nerve stimulation with observation of electromyograph…

Comparative analysis of fibrinolytic properties of Alteplase, Tenecteplase and Urokinase in an in vitro clot model of intracerebral haemorrhage.

Abstract Objective Hematoma lysis with recombinant tissue plasminogen activator (rtPA) has emerged as an alternative therapy for spontaneous intracerebral and intraventricular haemorrhage (ICH and IVH). However, the MISTIE III and CLEAR III trial failed to show significant improvement of favourable outcomes. Besides experimental and clinical trials revealed neurotoxic effects of rtPA. The demand for optimization of fibrinolytic therapy persists. Herein, we used our recently devised clot model of ICH to systematically analyse fibrinolytic properties of rtPA, tenecteplase and urokinase. Methods In vitro clots of human blood (size: 25 ml and 50 ml; age: 1.5 tenecteplase, 24 tenecteplase and 48…

Relation of early Photofrin uptake to photodynamically induced phototoxicity and changes of cell volume in different cell lines.

For efficacy of photodynamic therapy, selective uptake and retention of photoactive substances has been postulated. Therefore, measurements were performed to find out whether the photosensitiser Photofrin® is taken up differently in malignant and non-malignant cells in vitro . In addition, the sensitivity of malignant cells and nonmalignant cells to photodynamic exposure was investigated, by quantifying viability and volume alterations of the cells. Bovine aortic endothelial cells, mouse fibroblasts and amelanotic hamster melanoma cells were suspended in a specially designed incubation chamber under controlled conditions (e.g. pH, p O 2 , p CO 2 and temperature). After establishing constant…

Sauerstoffmetabolismus und intramuraler pH des Dünndarms bei mesenterialer Ischämie und Verhalten der Mikrozirkulation nach hyperton-hyperonkotischem Volumenersatz

Der akute Verschlus der A.mesenterica superior (AMS) fuhrt zu einer Ischamie des gesamten Dunndarms. Der Kollateralkreislauf kann mit unterschiedlichen Kompen-sationsmechanismen [2, 4] nur eine minimale und unzureichende Blutversorgung aufrechterhalten. Abhangig von der Ischamiedauer entsteht eine Gewebeschadigung. Bei der Reperfusion ischamischer Darmanteile werden zahlreiche vasoaktive Me-diatoren freigesetzt und eine Schocksituation verursacht, die den gesamten Orga-nismus beeintrachtigt und bei eingeschrankter Zirkulation eine Verlangerung der Darmischamie bewirkt. Die Tonometrie hat sich in einigen experimentellen Ver-suchsanordnungen als geeignete minimal-invasive Masnahme zur Abschat…

Does granulocyte-colony stimulating factor stimulate peripheral nerve regeneration? An experimental study on traumatic lesion of the sciatic nerve in rats.

Aim of the study. To analyse the therapeutic potential of granulocyte-colony stimulating factor (G-CSF) treatment using a rat model of traumatic sciatic nerve lesion. Clinical rationale for the study. G-CSF has proven strong neurotrophic properties in various models of ischaemic and traumatic brain injury. Fewer studies exist regarding the influence of G-CSF on posttraumatic peripheral nerve regeneration. Currently, the possibilities of pharmacological prevention or treatment of mechanical nerve injury are limited, and there is an urgent need to find new treatment strategies applicable in clinical situations. Material and methods . A controlled traumatic right sciatic nerve lesion was set u…

Early albumin infusion improves global and local hemodynamics and reduces inflammatory response in hemorrhagic shock.

Objective To evaluate the effects of an early, short-term albumin infusion on mesenteric microcirculation and global hemodynamics in hemorrhagic shock. Design A prospective, randomized study. Setting Animal laboratory at a university medical clinic. Subjects Seventeen Sprague-Dawley rats weighing 250–400 g. Interventions The rats underwent median laparotomy and exteriorization of an ileal loop for intravital microscopy of the mesenteric microcirculation. Volume-controlled hemorrhagic shock was provoked by arterial blood withdrawal (2.5 mL/100 g body weight for 60 mins), followed by a 4-hr reperfusion period. Albumin (20%) or 0.9% NaCl was administered intravenously as a continuous infusion …

Laser Doppler Scanning: How Many Measurements are Required to Assess Regional Cerebral Blood Flow?

This study was initiated to determine the optimal number of measuring sites necessary to estimate regional cerebral blood flow (CBF) under pathophysiological conditions.

Plasmavolumenverschiebung und Veränderungen der zentralen Hämodynamik nach Okklusion der A. mesent. sup. und anschließender Reperfusion

Im Schweinemodell wurde die Veranderung des Plasmavolumens bei verschiedenen therapeutischen Ansatzen des Reperfusionsschocks nach 2-stundiger Okklusion der A. mesent. sup. untersucht (keine Therapie, 5–50 μg Dopamin, 40 ml/kg KG isotone Reperfusion in 30 Min., 5 ml/kg KG 7,5% NaCl/10% HAES in 5 Min.). Der Plasmavolumenverlust von >30% in der Kontroll- und Dopamingruppe wurde nur durch die isotone Reperfusion vollstandig ausgeglichen. Die hyperton/hyperonkotische Therapie (-16% PV-Verlust) wies aber ein signifikant verbessertes HZV auf.

Using simple interrupted suture anastomoses may impair translatability of experimental rodent oesophageal surgery

Background/purpose: Irreproducibility and missing translatability are major drawbacks in experimental animal studies. Hand-sewn anastomoses in oesophageal surgery are usually continuous, whereas th...

Spreading Depression Induces Permanent Cell Swelling Under Penumbra Conditions

Background. Spreading depression (SD) is known to go along with temporary breakdown of ion gradients and cell swelling which spontaneously normalizes. Here, the effects of SD at reduced flow conditions as encountered in the ischemic penumbra are examined.

Cerebral blood flow alterations in a rat model of cerebral sinus thrombosis.

Outcome from sinus vein thrombosis is very variable, with symptoms from headache to coma. Experimental findings suggest that an involvement of cortical veins is necessary to affect the cerebral microcirculation. Laser Doppler flowmetry was used to investigate the regional and temporal changes in local cortical blood flow after experimental occlusion and thrombosis of the superior sagittal sinus and tributary cortical veins in rats. Thrombosis was induced by slow injection of kaolin-cephalin suspension after frontal and caudal ligation of the sagittal sinus in rats. Local cerebral blood flow was measured by laser Doppler flowmetry and correlated with parenchymal damage found 24 hours after i…

Towards a glioma model for surgical technique evaluation in the rat.

Evaluation of new surgical techniques in animal models is frequently challenging. This article describes the pitfalls, peculiarities and the final best applicable model for evaluating surgical techniques for glioma resection.The C6 glioma cell line and the Sprague-Dawley rat strain were selected. Fifty-thousand glioma cells were stereotactically transplanted in the left hemisphere of 137 male adult rats. Evaluation of solid tumour formation, tumour growth and scheduling of surgical resection was performed by MR scanning at 1, 2, and 4 weeks after transplantation and 3 and 6 months after tumour resection. Microsurgical tumour resection was performed with conventional techniques or with the w…

Nitric oxide modulates cerebral blood flow stimulation by acetazolamide in the rat cortex: a laser Doppler scanning study

Abstract The involvement of nitric oxide (NO) in cerebral blood flow (CBF) stimulation by acetazolamide was studied in anaesthetised, mechanically ventilated Wistar rats. CBF was monitored by laser Doppler scanning. Acetazolamide induced a long-lasting significant rCBF-increase. Application of N G -Nitro- l -arginine (L-NNA), an inhibitor of all NO synthetases (NOS), prevented CBF stimulation by acetazolamide. Continuous infusion of the exogenous NO donor SIN-1 (3-morpholinosydnonimine) suppressed L-NNA induced increases of mean arterial blood pressure without effect on rCBF in comparison to baseline. Additional acetazolamide injection then again caused a significant increase of rCBF in spi…

Significance of resting and stimulated cerebral blood flow for predicting the risk of hemodynamic cerebral ischemia in a model of chronic hemodynamic insufficiency.

OBJECTIVE: It has been postulated that patients with a compromised cerebrovascular reserve capacity (RC), defined as cerebral blood flow (CBF) response to acetazolamide (ACZ) (by percent), are at higher risk for ischemic stroke. The value of CBF and RC for predicting the risk of hemodynamically induced impairment of cerebral function is examined. METHODS: Both common carotid arteries were occluded in 22 Wistar-Kyoto rats. Thirty-one days later, mean arterial blood pressure was reduced to 40 mm Hg for 30 minutes. Laser Doppler scanning of CBF at resting conditions and after intraperitoneal administration of ACZ (0.1 mg/g body weight) was performed 30 minutes and 28 days after occlusion as we…

Flow and Pressure during Liver Preservation under ex situ and in situ Perfusion with University of Wisconsin Solution and Histidine-Tryptophan-Ketoglutarate Solution

Effective preservation of liver grafts is the first essential step for successful liver transplantation. Insufficient perfusion leads to ischemic-type biliary lesions after transplantation. Perfusion of the graft can be performed either in situ or ex situ, with gravity flow or pressure-controlled. Mainly University of Wisconsin (UW) and histidine-tryptophan-ketoglutarate (HTK) solutions are used widespread in clinical liver transplantation. Due to a persistent lack of data, we performed this systematic investigation of in situ and ex situ perfusion of liver grafts with HTK (low-viscous) and UW (high-viscous) solutions at different pressure steps on the perfusion solution (gravity flow, 50, …

Dynamic in vivo Imaging of Microvasculature and Perfusion by Miniaturized Confocal Laser Microscopy

<i>Introduction:</i> Microvasculature and associated pathologies mandate dynamic imaging. We evaluated a novel miniaturized confocal laser scanning probe for in vivo visualization of blood vessels, blood flow, cell tracking and perfusion in both healthy rodents and disease models.<i> Methods:</i> The hand-held confocal microscopy system allowed a 500- to 2,400-fold magnification at a dynamically variable imaging depth. Different intravital stains were used alone or in combination for tissue, nuclear, plasma and vascular endothelial cell staining and for blood flow visualization, and targeted staining for individual cell populations. <i>Results:</i> Precis…

Die Leukozytenakkumulation in der Dünndarmwand nach Okklusion der A. mesent. sup. und Therapie des Reper-fusionsschocks mit Dopamin, isotonem und hyperton-hyperonkotischem Volumenersatz

Die Leukozyten sind ein wichtiger Faktor in der Entstehung des Reperfusionsschadens. Der Nachweis der Leukozytenakkumulation erfolgt experimentell durch die Messung der leukozytenspezifischen Myeloperoxidase-Aktivitat in der Mucosa des Darms oder durch die direkte Beurteilung des intravaskularen Leukozytenverhaltens mit der intravitalen Mikroskopie, die vorwiegend an den Mesenterialgefasen durchgefuhrt wird. Neuere Studien zeigten jedoch, das die Leukozytenakkumulation nicht in allen Darmwandschichten identisch ist [1, 4].

Electrophysiology and neuronal integrity following systemic arterial hypotension in a rat model of unilateral carotid artery occlusion.

Patients with carotid artery stenosis may be particularly susceptible to hypotension-associated cerebral ischemia and subsequent neurological sequelae. Measuring somatosensory evoked potentials (SEP), electroencephalogram (EEG), direct current (DC) potential, and histology, we compared the temporal evolution of cortical functional perturbations as well as neuronal integrity in a model of unilateral carotid artery occlusion and systemic hypobaric hypotension (HH) at the lower limit of cerebral blood flow autoregulation (50 mm Hg). Serial measurements of EEG power spectra as well as SEP-amplitudes and latencies of N10.3 were performed before, during, and up to 60 min after 30 min-HH (n=7) or …

Control of brain temperature during experimental global ischemia in rats.

Temperature control during experimental ischemia continues to be of major interest. However, if exposure of brain tissue is necessary during the experiment, regional heat loss may occur even when the core temperature is maintained. Furthermore, valid non-invasive brain temperature monitoring is difficult in small rodents. This paper describes a method for both monitoring and maintenance of brain temperature during small animal preparations in a stereotaxic frame. The device used includes an ear-bar thermocouple probe and a small near-infrared radiator. The new equipment permitted to maintain peri-ischemic brain temperature at a desired level while carrying out non-invasive continuous record…

Tolerance-Inducing Dose of 3-Nitropropionic Acid Modulates bcl-2 and bax Balance in the Rat Brain: A Potential Mechanism of Chemical Preconditioning

Many studies have reported ischemia protection using various preconditioning techniques, including single dose 3-nitropropionic acid (3-NPA), a mitochondrial toxin. However, the cellular signal transduction cascades resulting in ischemic tolerance and the mechanisms involved in neuronal survival in the tolerant state still remain unclear. The current study investigated the mRNA and protein expression of the antiapoptotic bcl-2 and the proapoptotic bax, two antagonistic members of the bcl-2 gene family, in response to a single dose of 3-NPA, to global cerebral ischemia–reperfusion, and to the combination of both 3-NPA-pretreatment and subsequent global cerebral ischemia–reperfusion. Brain h…

An experimental study on magnetic esophageal compression anastomosis in piglets.

Abstract Introduction Fashioning a patent, watertight anastomosis in patients with esophageal atresia is a challenging task in pediatric surgery, particularly when performed under tension. A reproducible suture-less alternative would decrease operative time. We evaluated magnetic esophageal compression anastomoses in a novel bypass-loop swine model. Methods Eight-week-old piglets underwent thoracotomy to mobilize the esophagus at the carina to create a U-shaped loop. Custom-made 8 mm diameter Neodymium Magnets were inserted into the esophagus proximal and distal to the loop, then mated side-to-side at the future anastomosis site. Pigs were observed for 8 (n = 4), 10 (n = 6), and 12 (n = 2) …

Transcranial Doppler and Cortical Microcirculation at Increased Intracranial Pressure and during the Cushing Response

The effect of increased intracranial pressure on the flow velocity of the basilar artery was measured with transcranial ultrasonic Doppler in New Zealand White rabbits under alpha-chloralose anesthesia and artificial respiration. Laser Doppler flowmetry served to study changes of the cortical microcirculation. The results confirm a high inverse correlation of the diastolic flow velocity, the pulsatility index, and the resistance index with the cerebral perfusion pressure (CPP). During acute intracranial hypertension, however, these parameters do not show a good correlation with the local cortical blood flow. The absence of a correlation was evident over a wide CPP range down to values of 35…

Esophageal Biomechanics Revisited: A Tale of Tenacity, Anastomoses, and Suture Bite Lengths in Swine

Background Anastomotic tension has repeatedly been associated with anastomotic leakages after esophagectomy for cancer or esophageal atresia repair. We therefore aimed to determine which anastomotic technique would come as close as possible to the native esophagus in sustaining traction forces. Constant traction for several minutes at esophageal remnants and large suture bites are also considered relevant in long-gap esophageal atresia repair. Methods Porcine esophagi were subjected to linear traction using a motorized horizontal test stand. We compared breaking strengths of native esophagi to simple continuous, simple interrupted, stapled, and barbed suture anastomoses. We also investigate…

Comparative analysis of in situ versus ex situ perfusion on micro circulation in liver procurement--an experimental trial in a porcine model.

The Achilles heel of liver transplantation remains the biliary system. The crucial step for liver preservation is effective rinsing and perfusion of the peribiliary plexus (PBP). Due to the physiology of the vascular tree, it seems almost impossible to achieve the necessary physiologic ranges of pressure and flow by the in situ perfusion technique. We investigated the role of additional ex situ perfusion via the hepatic artery in this animal model.Fifteen German Landrace pigs underwent standardized multiorgan procurement. In situ perfusion and additional ex situ perfusion were performed consecutively. Meanwhile the external pressure applied to the perfusion system was increased stepwise. To…

Beyond Magnamosis: A Method to Test Sutureless Esophageal Anastomotic Devices in Living Swine by Creating an Esophageal Bypass Loop for Natural Oral Nutrition.

Abstract Introduction: Thoracoscopic esophageal atresia repair has become increasingly popular, but is still limited to a few expert centers and has some challenges and shortcomings. One of them ha...

In vitro and in vivo porphyrin accumulation by C6 glioma cells after exposure to 5-aminolevulinic acid

Several malignant tissues synthesize endogenous porphyrins after exposure to 5-aminolevulinic acid (5-ALA). The present experiments have been designed to elucidate whether the C6 glioma cell, a model cell for human malignant glioma, similarly synthesizes porphyrins when exposed to 5-ALA, and whether specific synthesis occurs when C6 cells are inoculated into rat brains to form a tumor. In this situation the blood-brain barrier may interfere with 5-ALA availability, and spreading of porphyrins with edema outside the tumor may occur. Flow cytometry is used to determine the course of cell volume and porphyrin fluorescence intensities in cultured C6 cells which are incubated in 1 mM 5-ALA. For …

Cecal Resection with Bipolar Sealing Devices in a Rat Model

Background: Bipolar sealing devices are routinely used to seal blood vessels. The aim of the study is to evaluate the feasibility and safety of colonic sealing with the use of the bipolar energy devices in rats as model for experimental appendectomy. Methods: Seventy-five male Wistar rats underwent a cecal resection with four different bipolar sealing devices or a linear stapler. The harvesting procedure was performed immediately or at postoperative day (POD) 7. The sealing front bursting pressure (BP) was measured in both groups. At POD7, the resection line was clinically examined and the hydroxyproline (HDP) levels were determined. Hematoxylin and Eosin (H&E) staining was used for histopa…

Cerebral embolic ischemia in rats: correlation of stroke severity and functional deficit as important outcome parameter.

The embolic MCA occlusion model in rats is used for recanalisation studies in acute stroke. In addition to the determination of lesion size, the assessment of functional outcome may improve the value of this model. Male Wistar rats were submitted to MCA clot embolism or sham surgery. In order to achieve a larger variety of lesion volume, 2 subgroups (each 7 animals) were subjected to differently sized emboli (30 and 40 mm). Follow-up period was 6 days. Outcome assessment consisted of a test battery including parallel bar crossing, observation of behaviour in an open field and an 8-arm maze and a neurological score with ten different sensorimotor and coordinative items. Animals were perfusio…

Biochemische Hirnprotektion

Local Cerebral Blood Flow in a Rat Cortical Vein Occlusion Model

The symptoms following sinus and vein occlusion observed in patients and experimental animals display a considerable variability that so far remains largely unexplained. In a rat cortical vein occlusion model using a photochemical thrombotic technique, we examined changes in the cerebral venous flow pattern by fluorescence angiography and regional cerebral blood flow (rCBF) and cerebral blood volume fraction (CBVF) by a modern laser Doppler “scanning” technique. Brain damage was assessed histologically. Fluorescence angiographic findings fell into two groups: group A, rats with an altered venous flow pattern after occlusion (n = 12), and group B, rats with interruption of blood flow and/or…

Microcirculatory alterations in a mongolian gerbil sinus-vein thrombosis model

Summary Background and purpose. The pathophysiology of sinus-vein thrombosis (SVT) is still controversial in patients and experimental animals, the microcirculatory alterations in particular. This study was designed to develop a new sinus-vein thrombosis model and to further elucidate pathophysiological events such as the relationship between local and regional cerebral blood flow and haemoglobin oxygen saturation (HbSO2), changes of the microvasculature, leukocyte behaviour and brain tissue damage. Methods. In a first experimental series, animals were divided into two groups which resulted from different procedures of inducing SVT. In the SSS middle occlusion group (SMO group), SVT was ind…

Implantable Sensors Based on Gold Nanoparticles for Continuous Long-Term Concentration Monitoring in the Body.

Implantable sensors continuously transmit information on vital values or biomarker concentrations in bodily fluids, enabling physicians to survey disease progression and monitor therapeutic success. However, currently available technologies still face difficulties with long-term operation and transferability to different analytes. We show the potential of a generalizable platform based on gold nanoparticles embedded in a hydrogel for long-term implanted biosensing. Using optical imaging and an intelligent sensor/reference-design, we assess the tissue concentration of kanamycin in anesthetized rats by interrogating our implanted sensor noninvasively through the skin. Combining a tissue-integ…

Motility analysis by means of video tracked markers

Abstract The motility of the gastrointestinal tract is crucial for digestive activity and dysfunction can lead to severe disease pattern. A method for analysing the motility is needed when treatment approaches shall be evaluated. Therefore markers attached to different locations on the stomach and the bowel of pigs are video tracked in this research study. The markers are designed to provide a high contrast and have an adhesive side for fixation. Above the operation field a video camera has been placed to film the markers during the procedure. To analyse the video data a special algorithm has been implemented. The algorithm requires a registration process at the beginning of each recording …

Measuring Intraoperative Oesophageal Anastomotic Tension With a Knot-Pusher–Mounted Dynamometer Is Not Quite Ready for Take-Off

Potent low dose platelet inhibitory effects of clopidogrel and aspirin on coronary thrombus formation in an animal model of acute unstable angina

SummaryApplication of clopidogrel before percutaneous coronary intervention in patients with acute coronary syndrome reduces the risk of cardiac events. Clopidogrel administration before surgery increases bleeding complications after CABG. Therefore, the antithrombotic effect of the low-dose combination of clopidogrel and aspirin was investigated in an in vivo pig model of coronary artery thrombus formation with cyclic flow reductions. The platelet inhibitory effect was determined by platelet aggregation and CFR, according to the methodology described by Folts. CFR were initiated by endothelial damage and placement of a constrictor around the LAD. 30 min after CFR were established, clopidog…

Application of C1-Esterase Inhibitor During Reperfusion of Ischemic Myocardium

Background—Complement activation during reperfusion of ischemic myocardium augments myocardial injury, and complement inhibition with C1-esterase inhibitor (C1-INH) at the time of reperfusion exerts marked cardioprotective effects in experimental studies. Application of C1-INH in newborns, however, was recently reported to have dangerous and even lethal side effects. This study addresses the essential role of dosage in studies using C1-INH.Methods and Results—Cardioprotection by C1-INH was examined in a pig model with 60 minutes of coronary occlusion followed by 120 minutes of reperfusion. C1-INH was administered intravenously 5 to 10 minutes before coronary reperfusion without heparin at a…

Surface Electromyography Reliably Records Electrophysiologically Evoked Internal Anal Sphincter Activity: A More Minimally Invasive Approach for Monitoring Extrinsic Innervation.

<b><i>Background:</i></b> Even in the case of minimally invasive pelvic surgery, sparing of the autonomic nerve supply is a prerequisite for maintaining anal sphincter function. Internal anal sphincter (IAS) innervation could be electrophysiologically identified based on processed electromyographic (EMG) recordings with conventional bipolar needle electrodes (NE). This experimental study aimed for the development of a minimally invasive approach via intra-anal surface EMG for recordings of evoked IAS activity. <b><i>Methods:</i></b> Six male pigs underwent nerve-sparing low anterior rectal resection. Electric autonomic nerve stimulations were …

Effects of hypertonic/hyperoncotic treatment and surgical evacuation after acute subdural hematoma in rats*

Objective: The treatment of acute subdural hematoma (ASDH) consists mainly of surgical evacuation of the hematoma. It is conceivable that early preoperative neuroprotection with hypertonic/hyperoncotic treatment (HHT) can improve survival rates. The present study investigated the benefit of treatment with hypertonic/hyperoncotic solution on functional and histologic outcome as supportive therapy accompanying surgical intervention. Design: Laboratory experiment. Setting: University laboratory. Subjects: Male Sprague-Dawley rats weighing 296–350 g (n = 56). Interventions: ASDH was induced through subdural infusion of 400 μL of autologous venous blood. Thirty minutes after subdural blood infus…

Experimentelle Untersuchungen zum Einsatz verschiedener atrialer Dipolkonfigurationen für die Vorhof-Stimulation mit VDD-Elektroden

Fourier-Analyse der P-Welle bei der VDD-Stimulation. Ein intra-individueller Vergleich verschiedener atrialer Dipolkonfigurationen

Der Frequenzinhalt der wahrgenommenen P-Welle wird bei der VDD-Stimulation sowohl von der Dipollokalisation, als auch von der Dipolkonfiguration beeinflust. Eine Vergroserung der Dipoloberflache resultiert in der Detektion tieferer Frequenzen.

Sex represents a relevant interaction in Sprague–Dawley rats: the example of oesophageal length*

Background: 8-week old Sprague Dawley rats represent the standard rodent model of oesophageal surgery, which is challenging and might be eased by larger oesophageal lengths. Therefore, we aimed to ...

Neuroprotection of S(+) ketamine isomer in global forebrain ischemia

The non-competitive N-methyl-D-aspartate (NMDA) receptor antagonist ketamine can block the action of excitotoxic amino acids in the central nervous system. S(+) ketamine has a 2-3 times higher anesthetic potency compared with the ketamine-racemate and also shows a higher neuroprotective efficacy in vitro. To determine the neuroprotective activity of S(+) ketamine compared with its R(-) stereoisomer in vivo, we examined the functional and neurohistological outcome in rats treated 15 min after global forebrain ischemia with S(+) ketamine in different dosages compared with R(-) ketamine. Influence of the treatment on regional cerebral blood flow (rCBF) and cortical oxygen saturation (HbO2) was…

In-vitro Untersuchungen zu photodynamisch induzierten Dosis-Wirkungs-Beziehungen

Im Gegensatz zu fruheren Annahmen wurde von Rodgers 1990 in Frage gestellt, das aus der Quantifizierung der Fluoreszenz im Gewebe Aussagen uber die Konzentration des Photosensibilisators noch uber die zu erwartenden photodynami-schen Effekte moglich sind (1).

EndothelinA receptor antagonist BSF-208075 causes immune modulation and neuroprotection after stroke in gerbils.

Leukocytes contribute to the ischemia-reperfusion injury. Recent studies suggested endothelins could be important mediators for leukocyte activation in stroke. We tested if the endothelinA receptor antagonist BSF-208075 (ambrisentan) could reduce an ischemic lesion by modulation of leukocyte-endothelium interactions. Twenty-four gerbils underwent either a sham operation (n=6) or 15 min of bilateral carotid artery occlusion resulting in global cerebral ischemia. Ischemic animals received normal saline (n=6), 5 mg/kg BSF-208075 (n=6) or 30 mg/kg (n=6) administered intravenously at 10 min of reperfusion. Leukocytes rolling or adhering to endothelium were counted by intravital microscopy in par…

Pringle maneuver increases the risk of anastomotic leakage after colonic resection in rats.

Abstract Background Many centers use the Pringle's maneuver during liver resections. Since this maneuver might impair healing of bowel anastomoses, we evaluated its influence on the healing of colonic anastomosis in rats. Methods Male Wistar rats underwent median laparotomy and sigmoid resection with end-to-end anastomosis under inhalation anesthesia. Thereafter, rats received a 25 minutes Pringle's maneuver (PM, group 1) or were kept under anesthesia for the same period of time (group 2). The anastomotic bursting pressure (BP) was measured on postoperative days (POD) 3, 6 and 9. Hematoxylin and Eosin (H&E) staining was used for histopathological evaluation of the anastomosis. The Mann-Whit…

Genomic response of the rat brain to global ischemia and reperfusion

To identify genes that are involved in ischemia response of the brain, we have evaluated changes of gene expression in rat cerebrum after 15 min complete global ischemia, followed by reperfusion for 1 h, 6 h or 24 h. The expression profiles of approximately 30,000 transcripts from three subjects in each group (including sham-operated controls) were monitored employing oligonucleotide microarrays. About 20,000 transcripts were detectable in rat brains. The levels of 576 transcripts (approximately 2.9%) were significantly altered in response to experimental ischemia. 419 transcripts were up- and 157 downregulated; 39 transcripts changed after 1 h reperfusion, 174 after 6 h and 462 after 24 h.…