Measure Of Tissue Damage Research Articles

A reproducible method for biochemical, histological and functional assessment of the effects of ischaemia\u2013reperfusion syndrome in the lower limbs

Current methodology described to mimic lower limb ischaemia–reperfusion injury (LL-IRI) does not accurately define the procedures and pressures exerted to induce and maintain ischaemia. In this piece of work, we propose a well-defined and detailed rat model that simulates the conditions established in clinical practice guidelines for tourniquet application and allows us to test treatments that aim to prevent/reduce LL-IRI. Eighty-six male WAG/RijHsd rats were subjected to hind limb IRI (LL-IRI), using a mechanical system applying a 1 kg tension to induce and maintain ischemia for 2 or 3 h, and assessed the damage caused by reperfusion at biochemical and muscular levels at different time points. At the biochemical level, both 2 and 3 h of ischemia induced changes (except for electrolyte levels); 3 h of ischemia induced greater changes in specific markers of muscular damage: creatine kinase (CK) and lactate dehydrogenase (LDH). At the histopathological level, 3 h of ischemia and 24 h of reperfusion was associated with an increase in hind limb girth, cross-sectional area, and weight and presence of neutrophils, as well as histological damage in more than 60% of muscle fibres. Our model allows to reliably reproduce the damage associated with the use of a pneumatic tourniquet. CK and LDH, as well as measures of tissue damage, allow to define and characterize the response to LL-IRI-related damage. A period of 3 h of ischemia followed by 3 h of reperfusion caused only local damage but showed greater sensitivity to detect differences in future studies on prophylactic treatments against LL-IRI.

Transcriptional modulations induced by proton irradiation in mice skin in function of adsorbed dose and distance

ABSTRACT Hadron therapy by proton beams represents an advanced anti-cancer strategy due to its highly localized dose deposition allowing a greater sparing of normal tissue and/or organs at risk compared to photon/electron radiotherapy. However, it is not clear to what extent non-targeted effects such as transcriptional modulations produced along the beamline may diffuse and impact the surrounding tissue. In this work, we analyze the transcriptome of proton-irradiated mouse skin and choose two biomarker genes to trace their modulation at different distances from the beam’s target and at different doses and times from irradiation to understand to what extent and how far it may propagate, using RNA-Seq and quantitative RT-PCR. In parallel, assessment of lipids alteration is performed by FTIR spectroscopy as a measure of tissue damage. Despite the observed high individual variability of expression, we can show evidence of transcriptional modulation of two biomarker genes at considerable distance from the beam’s target where a simulation system predicts a significantly lower adsorbed dose. The results are compatible with a model involving diffusion of transcripts or regulatory molecules from high dose irradiated cells to distant tissue’s portions adsorbing a much lower fraction of radiation.

The Effect of Shear Force on Skin Viability in Patients with Type 2 Diabetes.

Background Shear is a major risk factor in the development of diabetic foot ulcers, but its effect on the skin of patients with type 2 diabetes mellitus (DM) remains to be elucidated. The aim was to determine skin responses to shear in DM patients with and without diabetic polyneuropathy (DNP). Methods The forearm skin was loaded with 14.5 N shear (+2.4 kPa pressure) and with 3.5 kPa pressure for 30 minutes in 10 type 2 DM patients without DNP, 10 type 2 DM patients with DNP, and 10 healthy participants. A Sebutape collected IL-1α (measure of tissue damage). A laser Doppler flowmeter measured cutaneous blood cell flux (CBF) as a measure of the reactive hyperaemic skin response. Findings Reactive hyperaemia and IL-1α release was significantly increased after shear loading in all three groups and was higher compared to the responses to pressure loading. The reactive hyperaemic response after shear loading was impaired in patients with type 2 DM compared to healthy participants but did not differ between patients with and without DNP. The reactive hyperaemic response was negatively correlated with the blood glucose level but did not correlate with the DNP severity score. Interpretation Shear is important in the development of tissue damage, but the reparative responses to shear are impaired in patients with type 2 DM. DNP was not associated with altered skin responses, suggesting that the loss of protective sensation to sense shear to skin remains a key factor in the development of diabetic foot ulcers in patients with DNP.

Phase 2 Trial of Ibudilast in Progressive Multiple Sclerosis

BackgroundThere are limited treatments for progressive multiple sclerosis. Ibudilast inhibits several cyclic nucleotide phosphodiesterases, macrophage migration inhibitory factor, and toll-like receptor 4 and can cross the blood–brain barrier, with potential salutary effects in progressive multiple sclerosis.MethodsWe enrolled patients with primary or secondary progressive multiple sclerosis in a phase 2 randomized trial of oral ibudilast (≤100 mg daily) or placebo for 96 weeks. The primary efficacy end point was the rate of brain atrophy, as measured by the brain parenchymal fraction (brain size relative to the volume of the outer surface contour of the brain). Major secondary end points included the change in the pyramidal tracts on diffusion tensor imaging, the magnetization transfer ratio in normal-appearing brain tissue, the thickness of the retinal nerve-fiber layer, and cortical atrophy, all measures of tissue damage in multiple sclerosis.ResultsOf 255 patients who underwent randomization, 129 were assigned to ibudilast and 126 to placebo. A total of 53% of the patients in the ibudilast group and 52% of those in the placebo group had primary progressive disease; the others had secondary progressive disease. The rate of change in the brain parenchymal fraction was −0.0010 per year with ibudilast and −0.0019 per year with placebo (difference, 0.0009; 95% confidence interval, 0.00004 to 0.0017; P=0.04), which represents approximately 2.5 ml less brain-tissue loss with ibudilast over a period of 96 weeks. Adverse events with ibudilast included gastrointestinal symptoms, headache, and depression.ConclusionsIn a phase 2 trial involving patients with progressive multiple sclerosis, ibudilast was associated with slower progression of brain atrophy than placebo but was associated with higher rates of gastrointestinal side effects, headache, and depression. (Funded by the National Institute of Neurological Disorders and Stroke and others; NN102/SPRINT-MS ClinicalTrials.gov number, NCT01982942.)

A biopsychosocial understanding of lower back pain: Content analysis of online information.

(1) To develop a checklist to assess the representation of biopsychosocial lower back pain (LBP) online information; (2) to analyse publicly accessed online LBP information from a Google search for the degree that psychosocial contributors are described alongside the traditional biomedical approach to explaining LBP; (3) whether websites use information on pain biology to educate on LBP; (4) any inaccurate or false information regarding the mechanisms of LBP and; (5) the amount of websites certified by established benchmarks for quality health information. An online search was conducted using the Google search engines of six major English-speaking countries. Website content was analysed using three checklists developed for the purpose of this study - Biopsychosocial information categorisation checklist and scoring criteria; pain biology information checklist; and the inaccurate information checklist. Website quality was identified by the presence of an Health on the Net certification (HONcode). Of the fifteen websites analysed, the content of 26.7% of websites was classified as 'biomedical', 60% 'limited psychosocial' and 13.3% 'reasonable psychosocial'; 20% included information on pain biology; 46.7% inaccurately implied pain to be equal to tissue damage and 46.7% implied pathways specific to pain transmission; 40% were HONcode certified. Online LBP information retrieved through a Google search has limited to no integration of psychosocial or pain biology information. The focus on tissue pathology is further supported by the inaccurate descriptions of pain as equal to tissue damage and as an input to the central nervous system (CNS). Online LBP information needs to be guided by criteria more sensitive to the psychosocial contributors to pain. The online LBP information retrieved from a Google search needs to be guided by information more sensitive to the psychosocial contributors to pain and disability. This study also highlights the presence of inaccurate information that implied pain as a measure of tissue damage or as an input to the nervous system.

Application of Immunohistochemical Staining to Detect Antigen Destruction as a Measure of Tissue Damage

Electrocautery and directed energy devices (DEDs) such as lasers, which are used in surgery, result in tissue damage that cannot be readily detected by traditional histological methods, such as hematoxylin and eosin staining. Alternative staining methods, including 3-(4,5-dimethyl-thiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) to stain live tissue, have been reported. Despite providing superior detection of damaged tissue relative to the hematoxylin and eosin (H&E) method, the MTT method possesses a number of drawbacks, most notably that it must be carried out on live tissue samples. Herein, we report the development of a novel staining method, "antigen destruction immunohistochemistry" (ADI), which can be carried out on paraffin-embedded tissue. The ADI method takes advantage of epitope loss to define the area of tissue damage and provides many of the benefits of live tissue MTT staining without the drawbacks inherent to that method. In addition, the authors provide data to support the use of antibodies directed at a number of gene products for use in animal tissue for which there are no species-specific antibodies commercially available, as well as an example of a species-specific direct antibody. Data are provided that support the use of this method in many tissue models, as well as evidence that ADI is comparable to the live tissue MTT method.

Levels of phosphorylated axonal neurofilament subunit H (pNfH) are increased in acute ischemic stroke

For the study of stroke outcomes, there is the need for measurements of severity of stroke damage. Phosphorylated neurofilament heavy protein (pNfH) levels are elevated in axonal injury. We have measured levels of pNfH in stroke and correlated these levels with measures of stroke severity. Blood samples were collected from 54 ischaemic stroke patients at day 1, week 1 (days 7–10) and weeks 3–6, and an ELISA was used to measure pNfH levels in each patient at each time-point. Serum pNfH levels were significantly elevated in stroke patients compared to healthy controls. The levels were low at day 1, higher at day 7 and reached a peak at week 3, the latest day that we assessed. Significant associations were found between the pNfH levels at week 3 and early and stroke severity, size and outcome. Blood pNfH levels that reflect the severity of ischaemic stroke, are correlated with outcome and rise during the weeks after stroke. This may be a useful measure of tissue damage in stroke.

Aromatic- and di-carboxylates inhibit wound-induced phenolic accumulation in excised lettuce ( Lactuca sativa L.) leaf tissue

Excision of de-bladed 5-mm mid-rib segments from Romaine lettuce leaf tissue induced a five-fold rise in phenolic concentration when held at 10 °C for 48 h. Immersion in aqueous solutions of various di-carboxylates and aromatic carboxylates for 1 h reduced this wound-induced increase. The decrease was linear for di-carboxylates with increasing length from 3 to 14 carbons, with the phenolic concentration becoming less than the control for compounds with more than 6 carbons. Aromatic carboxylates with a hydroxyl at the 2 position and another at an even position produced an average reduction of 46%, while aromatic carboxylates with no hydroxyl groups, groups at odd positions, or adjacent groups produced an average 12% reduction. The decrease in wound-induced phenolic accumulation produced by 10 mM solutions of aromatic- and di-carboxylates was linearly correlated ( R 2 = 0.91) with the rate of carbon dioxide production, indicating possible tissue damage. This possibility was supported by the fact that most effective concentrations were highly correlated with increased ion leakage, another measure of tissue damage. While delaying the application of inhibitors of wound signal synthesis (e.g., n-alcohols) decreases their effectiveness, delaying the 1 h immersion in aromatic and di-carboxylate solutions for 4 h did not significantly reduce their effectiveness. Like mono-carboxylates, aromatic and di-carboxylates do not appear to be interfering with the synthesis or propagation of a wound signal, but appear to be inhibiting phenolic synthesis and/or accumulation at some subsequent step in the wound response.

Ischemia/reperfusion injury of skeletal muscle: Plasma taurine as a measure of tissue damage

Background. Cell membrane rupture by oxygen-derived free radicals is a systematic feature of ischemia/reperfusion (I/R) injury. High taurine concentration gradients in skeletal muscle prompted us to evaluate whether plasma taurine levels (pTau) are a useful marker of I/R injury after different periods of ischemia. Methods. Rabbits were randomly assigned to either 1 or 2.5 hours of hind-limb ischemia followed by 2 hours of reperfusion (groups IR1 [n = 12] and IR2.5 [n = 13], respectively). Corresponding sham groups (SHAM1 [n = 8] and SHAM2.5 [n = 9]) were used as controls. Analyzed parameters included histomorphometry and electron microscopy of skeletal muscle biopsies, pTau, and plasma level of malondialdehyde. Skeletal muscle function was assessed 3 weeks after I/R injury. Results. No significant morphologic changes were detectable at the end of ischemia. After reperfusion, mild interstitial edema with intact muscle cell membranes developed in IR1 group; pTau was not increased. IR2.5 group, by contrast, showed severe interstitial edema formation (interfiber area increased by 112%, P <.005), microvascular constriction (microvessel area decreased by 33%, P <.0005), and damage to the muscle cell membranes that was confirmed by the increased plasma malondialdehyde. pTau was higher than in the SHAM2.5 group (P <.0005). Pronounced cell damage in IR2.5 group resulted in impaired muscle function (maximal tetanic tension was reduced 2 times, P <.005) but not in IR1 group. Conclusion. Skeletal muscle tolerates 1 h/2 h but not 2.5 h/2 h of I/R, the latter resulting in interstitial edema formation, microvascular constriction, and a late muscle dysfunction. Cell membrane rupture through stimulated lipid peroxidation promotes leakage of intracellular taurine, leading to increased pTau after reperfusion and may be considered as prognostically unfavorable in terms of organ function reversibility. In the rabbit model, pTau seems to be a sensitive marker of I/R injury to skeletal muscle. (Surgery 2003;133:91-100.)

Correlations between clinical and MRI involvement in multiple sclerosis: assessment using T1, T2 and MT histograms

The degree of disability and cerebellar and brainstem impairments in multiple sclerosis (MS) patients were correlated with several magnetic resonance imaging (MRI) measures of tissue damage in the whole brain, cerebellum and brainstem to determine the relative contributions of the factors underlying the development of disability in MS. Dual-echo conventional spin-echo, T(1)-weighted and magnetization transfer (MT) scans were obtained from 72 patients with MS and 20 age- and sex-matched controls. The following MRI-derived quantities were considered for the brain as a whole, for the cerebellum and for the brainstem: (a) the number and volume of lesions seen on T(2)-weighted images; (b) the number and volume of lesions seen on T(1)-weighted images; (c) the size of these structures measured on T(1)-weighted scans; (d) the average MT ratio (MTR), peak height and peak position for the MT histogram. With univariate analysis, many MRI measures were significantly different in patients with different levels of disability or cerebellar and brainstem functional system impairments. However, with multivariate analysis, only whole-brain average MTR was significantly related to physical disability, while cerebellar and brainstem T(1) lesion volume and average MTR were related to cerebellar and brainstem impairment. This study shows that increased pathological damage in clinically eloquent sites is the major cause of disability in patients with MS. It also suggests that measures derived from MT histogram analysis and T(1) hypointense lesion load should be considered when evaluating long-term MS evolution.

Effect of free-radical scavengers and hyperbaric oxygen on random-pattern skin flaps.

To determine whether treatment with the combination of hyperbaric oxygen and free-radical scavengers or inhibitors would result in increased skin-flap survival. An animal model with male Sprague-Dawley rats was used. The flap was a cranial-based dorsal 3 x 12-cm random-pattern skin flap that included the panniculus carnosus. Rats were randomly assigned to one of 10 treatment groups. The radical scavengers superoxide dismutase, catalase, and alpha-tocopherol acetate and the inhibitor allopurinol were used to combat or scavenge radicals. Oxygen (100%) treatments were for 90 minutes at 2.5 atm absolute daily. At 7 days, the flaps were examined for survival by fluorescein injection. Lipid peroxidation as a measure of tissue damage was measured by thiobarbituric acid-malondialdehyde analysis. The combination of treatments resulted in significantly increased flap survival compared with untreated controls (P < .05) except in the group treated with allopurinol and hyperbaric oxygen. Lipid peroxidation was inhibited by the superoxide dismutase plus catalase and the alpha-tocopherol treatments but not by treatment with allopurinol. Moderate doses of radical scavengers or antioxidants coupled with a conservative hyperoxic exposure regimen can result in the increased survival of random-pattern skin flaps.

The effect of force magnitude on extractable bone resorptive activity and cemental cratering in orthodontic tooth movement.

The delay period in the orthodontic tooth movement cycle was studied with respect to the stimulation of an extractable bone resorptive activity and the degree of tissue damage at sites of greatest pressure. Total area of cemental cratering was used as a measure of tissue damage. Both bone resorptive activity and tissue damage were found to be significantly increased with moderate and high forces, a fact which supports the suggestion that bone resorptive activity is related to tissue damage.

Effect of methylprednisolone upon technetium-99m pyrophosphate assessment myocardial necrosis in the canine countershock model.

Repeat DC countershock reproducibly results in myocardial necrosis in dogs. In this model, myocardial technetium-99m pyrophosphate (PYP) uptake correlates linearly with tissue creatine kinase depletion (r = -0.83). The effect of pretreatment with methylprednisolone (MP) was studied with PYP in 25 dogs. In myocardium damaged by countershock, 12 MP dogs had higher tissue radioactivity sample:normal (S:N) ratios than control (P less than 0.05), suggesting increased tissue injury. However, by several other measures of tissue damage, the two groups did not differ. MP-elevated PYP S:N ratios were explained by reduced PYP activity in normal myocardium of MP dogs. Further experiments in 21 dogs revealed that renal PYP clearance, which correlated with glomerular filtration rate (GFR) as measured by creatinine clearance, was increased in Mp dogs, resulting in accelerated urinary excretion of PYP (46.9+/-3.6 vs 35.8+/-2.4 percent injected dose in one hour, P less than 0.01), and reduced blood PYP. Thus MP does not modify countershock-induced myocardial injury. However, by increasing GFR, MP increased PYP excretion, resulting in lowered blood and normal zone myocardial PYP, thereby spuriously affecting myocardial PYP tissue uptake data.