A dual marker technique to estimate individual feed intake in young pigs.

Accurate estimation of individual feed intake (FI) of pigs could help better understand the variation in performance between individual animals. We studied dual marker methods to estimate individual FI in pigs. This method is based on the measurement of the ratio between two indigestible markers in faeces. Twelve 6.5-week-old individually housed male pigs were assigned to one of three oral dosing treatments supplying 180 mg of ytterbium chloride (YbCl3)/day and 111 mg of dotriacontane (C32)/day as reference markers, either once (R1), three times (R3) or five times (R5) daily. Pigs were offered a diet containing 0.46 g/kg of chromium chloride (CrCl3) and 0.15 g/kg of hexatriacontane (C36) as in-feed markers. The experiment lasted for 10 days: days -5 to 0: adaptation; days 1-3: dosing of reference marker; days 2-4: total faecal collection. Spot faecal samples were taken on day 3 at 1200 h, 1700 h and on day 4 at 0700 h. Pigs were fed restrictedly three times daily, at 133.6 g/kg BW0.60. Individual measured FI was recorded daily and was compared to predicted FI using the ratio of the dual marker pairs (Yb:Cr and C32:C36), both in total faecal collection and spot samples. Due to unequal variance, R1 pigs were omitted from the statistical treatment comparison. When using total faecal collection samples, the absolute prediction error (APE) (predicted FI minus measured FI) in R3 and R5 pigs was numerically lower than in R1 pigs, regardless of the marker pair used. The APE measured by C32:C36 was numerically lower than measured by Yb:Cr at all frequencies, and significantly (P = 0.039) in R3 pigs (C32:C36: 0.15 ± 0.02 kg/day; Yb:Cr: 0.29 ± 0.04 kg/day). This was related to a larger difference in faecal recovery between Yb and Cr compared with C32 and C36. Daily total faecal collection revealed that for R3 pigs, starting faecal collections 2 days after the onset of provision of the reference marker improved the APE when compared with starting after 1 day. When using C32:C36 to predict feed intake, pooled, but not single spot samples gave similar APEs compared with total faecal collections. Therefore, we recommend dosing the reference marker three times per day for 2 days on days 1 and 2, combined with pooled spot faecal sampling collected on days 3 and 4. In this way, absolute prediction errors of 10%-15% of simultaneously measured intakes of multiple nutrient resources in a complex housing system are feasible using the dual marker technique.

Effect of rubber flooring on group-housed sows' gait and claw and skin lesions.

This study evaluated the influence of floor type on sow welfare in terms of lameness, claw lesions, and skin lesions. In a 2 × 3 factorial design, we have investigated the effect of rubber coverings on concrete floors and the effect of 3 levels of dietary zinc supplementation on locomotion and claw and skin lesions in group-housed sows. Six groups of 21 ± 4 hybrid sows were monitored during 3 successive reproductive cycles. The sows were group housed from d 28 after insemination (d 0) until 1 wk before expected farrowing date (d 108) in pens with either exposed concrete floors or concrete floors covered with rubber in part of the lying area and the fully slatted area. During each reproductive cycle, locomotion and skin lesions were assessed 4 times (d 28, 50, 108, and 140) and claw lesions were assessed twice (d 50 and 140). Results are given as least squares means ± SE. Locomotion and claw scores were given in millimeters, on analog scales of 150 and 160 mm, respectively. Here, we report on the effect of floor type, which did not interact with dietary zinc concentration ( > 0.10 for all variables). At move to group (d 28) and mid gestation (d 50), no differences between floor treatments were seen in locomotion ( > 0.10). At the end of gestation (d 108), sows housed on rubber flooring scored 9.9 ± 4.1 mm better on gait ( < 0.001). Regarding claw disorders, both parameters "heel overgrowth and erosion" (difference of 4.6 ± 1.8 mm; = 0.01) and "heel-sole crack" (difference of 3.1 ± 1.5 mm; = 0.04) scores were better for sows on rubber flooring at mid gestation (d 50). However, sows on rubber flooring scored worse for "vertical cracks in the wall horn" (difference of 3.4 ± 1.7 mm; = 0.04). At the end of lactation (d 140), both "white line" (difference of 2.9 ± 1 mm; = 0.02) and "claw length" (difference of 4.7 ± 1.4 mm; < 0.001) had better scores on rubber flooring. No differences for skin lesions were observed between floor treatments. The improved scores for gait toward the end of gestation and some types of claw disorders at mid gestation suggest that rubber flooring in group housing has a beneficial effect on the overall leg health of sows. The documented increase in vertical cracks in the wall horn at d 50 requires further investigation.

The burned ear; possibilities and challenges in framework reconstruction and coverage.

Ear reconstruction remains a challenging procedure, especially in burn victims. The ear is particularly vulnerable to thermal injury because of its location and the thin integument. The thermal injury could subsequently include skin and the deeper located auricular cartilage framework. This type of injury could have long lasting mutilating effect not only because the ear's morphology is mainly related to this framework but also because it will not recover or regenerate once injured. Grafts of costal cartilage or synthetic materials might replace missing cartilage. However, the poor quality of the adjacent skin and subcutaneous tissues makes the reconstruction of a burned ear an even more daunting procedure than congenital or many oncologic indications. As such, regeneration of the skin will be the next step in reconstruction of the burned ear. There is still much development and research to be done, but encouraging results have been shown in tissue engineering of skin and cartilage. Furthermore, 3D (bio)printing of cartilage to facilitate reproduction of the ear's complex shape certainly has potential and might find an interesting role in ear reconstruction. In this review, different clinical challenges and options for ear reconstruction in burn patients are described. Subsequently, although still far from large scale clinical application, state of the art developments in the field of tissue engineering and 3D (bio)printing are also discussed.

Effect of locomotion score on sows' performances in a feed reward collection test.

Sows housed in groups have to move through their pen to fulfil their behavioural and physiological needs such as feeding and resting. In addition to causing pain and discomfort, lameness may restrict the ability of sows to fulfil such needs. The aim of our study was to investigate the extent to which the mobility of sows is affected by different degrees of lameness. Mobility was measured as the sow's willingness or capability to cover distances. Feed-restricted hybrid sows with different gait scores were subjected to a feed reward collection test in which they had to walk distances to obtain subsequent rewards. In all, 29 group-housed sows at similar gestation stage (day 96.6 ± 7 s.d.) were visually recorded for gait and classified as non-lame, mildly lame, moderately lame or severely lame. All sows received 2.6 kg of standard commercial gestation feed per day. The test arena consisted of two feeding locations separated from each other by a Y-shaped middle barrier. Feed rewards were presented at the two feeders in turn, using both light and sound cues to signal the availability of a new feed reward. Sows were individually trained during 5 non-consecutive days for 10 min/day with increasing barrier length (range: 0 to 3.5 m) each day. After training, sows were individually tested once per day on 3 non-consecutive days with the maximum barrier length such that they had to cover 9.3 m to walk from one feeder to the other. The outcome variable was the number of rewards collected in a 15-min time span. Non-lame and mildly lame sows obtained more rewards than moderately lame and severely lame sows (P<0.01). However, no significant difference was found between non-lame and mildly lame sows (P=0.69), nor between moderately lame and severely lame sows (P=1.00). This feed reward collection test indicates that both moderately lame and severely lame sows are limited in their combined ability and willingness to walk, but did not reveal an effect of mild lameness on mobility. These findings suggest that moderately and more severely lame sows, but not mildly lame sows, might suffer from reduced access to valuable resources in group housing systems.

Ischemic cardiomyopathy and cerebral infarction in a young patient associated with khat chewing.

Khat is a stimulating agent used by many people in the Horn of Africa and the Arabian peninsula. Khat chewing is a known cardiovascular risk factor and is thought to cause vasoconstriction, systemic hypertension, and thrombogenicity. A 33-year-old Somalian man initially presented with loss of neurological function of the left arm, hazy vision, and headache. He smokes tobacco and chews two bundles of khat a week for more than 10 years. His ECG on admission showed a Q wave in V1 and V2 and 2 mm ST-elevations in V1, V2, and V3 and a terminal negative T wave in I, aVL, V2, V3, and V4, consistent with a recent, evolving anterior infarction. A noncontrast enhanced CT of the brain showed ischemia in the right middle cerebral artery vascular territory. An MRI showed recent ischemia in the vascular territory of the posterior division of the right middle cerebral artery. Coronary angiography showed a 70% stenosis with haziness of the proximal left anterior descending artery. Diagnostic tests and imaging are consistent with recent myocardial infarction in the LAD vascular territory because of coronary spasm and cerebral infarction in the middle cerebral artery vascular territory probably related to khat chewing.

Developing a parametric ear model for auricular reconstruction: a new step towards patient-specific implants.

INTRODUCTION: Ear reconstruction is a tedious and demanding surgical procedure and the implant framework used is essential for the esthetic result. The outcome of a reconstructed ear, however, is not necessarily limited to the implant shape but rather to the available options of transplantable tissue for coverage. Apart from the visual aesthetics, ear reconstruction subsequently also requires implant dimensions to be adapted to the surgical possibilities. In this article, we have brought different disciplines together to develop a customizable ear model for 3D printing of ear implants. MATERIAL AND METHODS: Computed tomography (CT) scans were made of 4 human cadaver ears before and after soft tissue dissection using a Discovery 750 High Definition Freedom Edition scanner (GE, Milwaukee, WI, USA) and subsequently converted into an STL data set using Mimics Software (Materialise, Leuven, Belgium). These scans were then used to develop a fully adjustable parametric model based on the essential ear anatomy using Rhinoceros and Grasshopper software. RESULTS: To determine the quality of the developed models, directed Hausdorff distance (DHD) was applied as the basis for measuring the similarity between the parametric model and the ear cartilage scanning data. Two methods were used. The mean directed Haussdorff distance (MDHD) was calculated based on the distribution of point sets showing an average similarity of 0.8 mm (±0.05 mm). The mean similarity coefficient (SC) of the model and scan surfaces was 94% with a 2-mm threshold. CONCLUSION: This study shows that a parametric standard model could be used as a feasible method to generate custom implants based on existing ear images.

Tissue engineering in burn scar reconstruction.

Nowadays, most patients with severe burns will survive their injury. This evolution is accompanied by the challenge to cover a large percentage of total body surface area burned. Consequently, more and more patients have to deal with the sequelae of burn scars and require (multiple) reconstructions. This review provides a gross overview of developments in the field of tissue engineering for permanent burn wound coverage and reconstructive burn surgery, focusing on usage and clinical effectiveness. Not only skin substitutes will be discussed but also the replacement of subcutaneous fat tissue and cartilage.

Right-to-left atrial shunting with normal intracardiac pressures following cardiac surgery: pathophysiology and management.

We present a case of acute respiratory insufficiency with right-to-left atrial shunting under normal intracardiac pressures discovered several days after aortic surgery for aortic dissection. We discuss the possible mechanisms and management of right-to-left atrial shunting through an atrial septum defect with normal intracardiac pressures following cardiac surgery.

Cell transplantation for cardiac regeneration: where do we stand?

During the last decade transplantation of cells into the heart has emerged as a novel therapy for the prevention and treatment of heart failure. Although various cell types have been used, most experience has been obtained with the progenitor cells of skeletal muscle, also called myoblasts, and a wide array of bone marrow-derived cell types. The first preclinical studies demonstrated an improvement in global and regional heart function that was attributed mainly to a direct contractile effect of the transplanted cells. Furthermore, it was suggested that multiple cell types are able to form true cardiomyocytes and truly 'regenerate' the myocardium. More recent studies have questioned these early findings. Other mechanisms such as paracrine effects on the infarct and remote myocardium, a reduction in adverse remodelling and improvement of mechanical properties of the infarct tissue likely play a more important role. On the basis of encouraging preclinical studies, multiple early-phase clinical trials and several randomised controlled trials have been conducted that have demonstrated the feasibility, safety and potential efficacy of this novel therapy in humans. This review summarises the available evidence on cardiac cell transplantation and provides an outlook on future preclinical and clinical research that has to fill in the remaining gaps. (Neth Heart J 2008;16:88-95.).

Reduction in infarct size, but no functional improvement after bone marrow cell administration in a porcine model of reperfused myocardial infarction.

AIMS: Stem cell therapy after myocardial infarction (MI) has been studied in models of permanent coronary occlusion. We studied the effect of intracoronary administration of unselected bone marrow (BM) and mononuclear cells (MNC) in a porcine model of reperfused MI. METHODS AND RESULTS: In 34 swine, the left circumflex coronary artery was balloon-occluded for 2 h followed by reperfusion. Ten swine without MI served as controls. All swine underwent magnetic resonance imaging (MRI) 1 week post-MI. The next day, 10 of the 30 surviving MI swine received BM, 10 other MI swine received MNC, and the remaining MI swine received medium intracoronary. Four weeks later, all swine underwent a follow-up MRI. One week after MI, end-diastolic volume (92+/-16 mL) and left ventricular (LV) weight (78+/-12 g) were greater, whereas ejection fraction (40+/-8%) was lower than in controls (69+/-11 mL, 62+/-13 g, and 53+/-6%). Injection of BM or MNC had no effect on the MI-induced changes in global or regional LV-function. However, there was a significant reduction in infarct size 4 weeks after MNC injection (-6+/-3%) compared with the medium (-3+/-5%). CONCLUSION: Intracoronary injection of BM or MNC in swine does not improve regional or global LV-function 4 weeks after injection. However, a reduction in infarct-size was noted after MNC injection.

In vitro imaging of single living human umbilical vein endothelial cells with a clinical 3.0-T MRI scanner.

Iron oxide-labelled, single, living human umbilical vein endothelial cells (HUVECs) were imaged over time in vitro using a clinical 3.0-T magnetic resonance (MR) microscopy system. Labelling efficiency, toxicity, cell viability, proliferation and differentiation were assessed using flow cytometry, magnetic cell sorting and a phenanthroline assay. MR images were compared with normal light and fluorescence microscopy. Efficient uptake of iron oxide into HUVECs was shown, although with higher label uptake dose-dependent cytotoxic effects were observed, affecting cell viability. For MR imaging, a T2* weighted three-dimensional protocol was used with in-plane resolution of 39 x 48 microm2 and 100-microm slices with a scan time of 13 min. MRI could detect living cells in standard culture dishes at single-cell resolution, although label loss was observed that corresponded with the intracellular iron measurements. MR microscopy using iron oxide labels is a promising tool for studying HUVEC migration and cell biology in vitro and in vivo, but possible toxic effects of label uptake and loss of label over time should be taken into account.

High-resolution magnetic resonance imaging of iron-labeled myoblasts using a standard 1.5-T clinical scanner.

Myoblast transplantation is a promising means of restoring cardiac function in infarcted areas. For optimization of transplant protocols, tracking the location and fate of the injected cells is necessary. An attractive imaging modality for this is magnetic resonance imaging (MRI) as it is noninvasive and as iron-labeled myoblasts provide a signal attenuation in T2*-weighted protocols. The aim of this study was to develop an efficient iron-labeling protocol for myoblasts and to visualize single-labeled cells using a clinical 1.5-T scanner. Pig myoblasts were labeled with a superparamagnetic iron oxide (SPIO) agent using a liposome transfection agent. Labeling efficiency, toxicity, cell viability, and proliferative capacity were measured for 10 days. Magnetic resonance (MR) of myoblast cultures used a T2*-weighted three-dimensional protocol with a maximum in-plane resolution of 19.5 x 26.0 microm2 and 50 microm slices. Use of liposomes improved SPIO labeling efficiency. Labeling did not induce toxicity or affect cell viability or proliferation. The cell distribution as observed with light and fluorescence microscopy matched the signal voids observed in the MRI datasets. Liposomes promote fast, nontoxic and efficient SPIO labeling of myoblasts that can be tracked by MRI microscopy in clinical scanners using susceptibility-weighted protocols.

Cardiac transplantation of skeletal myoblasts for heart failure.

Heart failure has become the most prevalent cardiovascular syndrome, and its incidence continues to increase. Most cases of heart failure develop as a result of myocardial infarction. Although current treatment modalities have brought us the opportunity to reduce mortality and morbidity after myocardial infarction, our progress has plateaued due to our inability to treat the underlying problem, death of cardiomyocytes. Recently, a new option has emerged. Transplantation of undifferentiated cells into the damaged heart is a promising new treatment modality. These cells may have the capability of adapting to the cardiac environment, regenerating the damaged muscle, restoring cardiac function and preventing transition to heart failure. During the last few years many cell types have been proposed for cardiac repair and promising pre-clinical studies have moved some of these into the clinic. The most widely studied cell type is the progenitor cell of adult muscle, or the myoblast. When transplanted into the heart myoblasts are able to engraft and to a large degree regenerate the infarcted area. Although the feasibility of myoblast transplantation has been proven in animal models of infarction, many questions remain unanswered. In this review we will try to present an overview of where intracardiac myoblast transplantation stands and where it is heading. We also provide our insight into the future potential for myoblast transplantation clinically.

Comparison of E-selectin expression at mRNA and protein levels in murine models of inflammation.

BACKGROUND: Drug targeting to activated endothelial cells via E-selectin is currently being explored as a new approach to treat chronic inflammatory disorders. This approach uses E-selectin directed antibodies as carrier molecules to selectively deliver anti-inflammatory drugs into activated endothelial cells, thereby theoretically decreasing drug-associated side-effects. Therapeutic effects of developed drug targeting constructs will have to be tested in animal models of inflammation, in which E-selectin is expressed during the course of the disease. In this study several murine models of inflammation were investigated regarding expression of E-selectin. METHODS: E-selectin expression was determined both at the mRNA level using RT-PCR and at the protein level by immunohistochemistry using two monoclonal antibodies (10E9.6 and MES-1). The models studied included delayed type hypersensitivity induced skin inflammation, dextran sodium sulphate induced colitis, kidney ischemia/reperfusion injury, atherosclerosis in ApoE knockout mice, and collagen induced arthritis. RESULTS: In all animal models E-selectin mRNA expression was detected, although to a different extent. In contrast, only the delayed type hypersensitivity model and, to a minor extent, the collagen induced arthritis model showed E-selectin protein expression. CONCLUSION: These results stress the need to determine E-selectin protein expression and not only mRNA expression, when choosing an animal model for testing E-selectin directed drug targeting preparations. In addition, in the arthritis model, E-selectin protein detection was dependent on the particular anti-E-selectin antibody used. This finding may not only have implications for the development and/or choice of homing devices to be used in E-selectin directed drug targeting preparations, but also for inflammation research in general.