Ischaemia and reperfusion effects on skeletal muscle tissue: morphological and histochemical studies

This was a study on the oxidative stress due to ischaemia (I) and reperfusion (R) in skeletal muscle tissue. Using a tourniquet, groups of rats were submitted to ischaemia for 4 h, followed by different reperfusion periods. The animals were divided in four groups: control; 4 h of ischaemia (IR); 4 h of ischaemia plus 1 h reperfusion (IR-1 h); 4 h of ischaemia plus 24 h reperfusion (IR-24 h); and 4 h of ischaemia plus 72 h reperfusion (IR-72 h). At the end of the procedures, samples of soleus muscle were collected and frozen in n-hexane at -70 degrees C. Cryostat sections were submitted to haematoxylin-eosin, succinate dehydrogenase (SDH) and nicotinamide adenine dinucleotide-tetrazolium reductase (NADH-TR) stains. An additional muscle sample was processed for electron microscopy. No alterations were found in control animals. IR group showed fibres had normal aspect besides some round, acidophilic and hypertrophic fibres. There were several fibres with angular outlines and smaller diameters in this group compared with control group. NADH-TR/SDH reaction was moderately intense in most fibres. In some fibres, cytoplasm showed areas without activity and other fibres had very intense reactivity. IR-1 h group showed oedema hypercontracted fibres with disorganized myofibrils, mitochondria with focal lesions and dilated sarcoplasmic reticulum. NADH-TR/SDH reaction was moderate to weak. IR-24 h showed intense inflammatory infiltrate in the endomysium and perimysium. NADH-TR/SDH reaction was similar to IR-1 h. IR-72 h showed necrotic fibres, areas with inflammatory infiltrate, reduced muscle fibres at different stages of necrosis and phagocytosis, and many small round and basophilic fibres characterizing a regeneration process. NADH-TR/SDH reaction was weak to negative. Our results suggest that ischaemia and the subsequent 1-, 24- and 72-h reperfusions induced progressive histological damage. Although progressive, it may be reversible because there were ultrastructural signs of recovery after 72-h reperfusion. This recovery could in part be due to the low oxidative stress identified by the morphological and histochemical analysis.

Effect of allopurinol on the kidney function, histology and injury biomarker (NGAL, IL 18) levels in uninephrectomised rats subjected to ischaemia-reperfusion injury

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Ischaemia modified albumin in horses exercised at different intensities in treadmill

Ischaemia modified albumin (IMA) is considered a biomarker of myocardial ischaemia in humans in contrast to other biomarkers released when cardiac necrosis occurs. Little is known about release conditions of IMA in exercise and this is the first report in equine species. For this purpose, ten clinically healthy untrained horses were submitted to a high intensity test (HIT) followed by a low intensity test (LIT) seven days later. Blood samples were taken before, during and immediately after exercise, and 15 min and 30 min thereafter. Serum IMA, lactate and albumin, and plasma malondialdehyde (MDA) were determined. There were no significant changes in IMA concentration in any of the exercise tests. There was also a negative correlation between IMA and albumin levels in both tests, and between IMA and lactate levels in LIT, suggesting possible assay interferences. It was concluded that HIT and LIT did not promote significant changes in IMA concentration in horses under these conditions.

Neoangiogênese de retalhos cutâneos em ratos tratados com óleo de copaíba

O objetivo deste trabalho foi avaliar morfometricamente a neoangiogênese de retalhos cutâneos subdérmicos em ratos tratados com óleo de copaíba (Copaifera langsdorffii) em pomada a 10%. O delineamento experimental foi inteiramente casualizado, composto por três recursos farmacológicos. Foram utilizadas dez repetições para cada recurso, e cada animal foi considerado uma unidade experimental. Trinta ratos Wistar foram submetidos à elevação do retalho cutâneo dorsal subdérmico e distribuídos em três grupos: grupo controle absoluto, no qual os animais não receberam nenhum tratamento; grupo controle, no qual os animais receberam tratamento tópico diário com pomada com apenas veículo (glicerina e vaselina); e grupo tratado, no qual os animais foram tratados diariamente com óleo de copaíba em pomada a 10%. Os ratos foram tratados e observados por oito dias após o ato operatório. No oitavo dia de pós-operatório, realizou-se a análise macroscópica do retalho e foram coletados fragmentos das porções cranial, média e caudal do retalho cutâneo para análise histopatológica. A análise morfométrica mostrou diferença significativa para o número de novos vasos sanguíneos nas partes média e caudal do retalho cutâneo no grupo tratado. O óleo de copaíba mostra-se eficiente no aumento da neoangiogênese em retalhos cutâneos subdérmicos de ratos.

Intra-articular pressure profiles of the cadaveric equine fetlock joint in motion

The study of the influence of motion and initial intra-articular pressure (IAP) on intra-articular pressure profiles in equine cadaver metatarsophalangeal (MTP) joints was undertaken as a prelude to in vivo studies, Eleven equine cadaver MTP joints were submitted to 2 motion frequencies of 5 and 10 cycles/min of flexion and extension, simulating the condition of lower and higher (double) rates of passive motion. These frequencies were applied and pressure profiles generated with initial normal intra-articular pressure (-5 mmHg) and subsequently 30 mmHg intra-articular pressure obtained by injection of previously harvested synovial fluid.The 4 trials performed were 1) normal IAP; 5 cyles/min; 2) normal IAP; 10 cycles/min; 3) IAP at 30 mmHg; 5 cycles/min and 4) IAP at 30 mmHg; 10 cycles/min. The range of joint motion applied (mean +/- s.e.) was 67.6 +/- 1.61 degrees with an excursion from 12.2 +/- 1.2 degrees in extension to 56.2 +/- 2.6 degrees in flexion, Mean pressure recorded in mmHg for the first and last min of each trial, respectively, were 1) -5.7 +/- 0.9 and -6.3 +/- 1.1; 2) -5.3 +/- 1.1 and -6.2 +/- 1.1; 3) 58.8 +/- 8.0 and 42.3 +/- 7.2; 4) 56.6 +/- 3.7 and 40.3 +/- 4.6. Statistical analyses showed a trend for difference between the values for the first and last minute in trial 3 (0.05>P<0.1) with P = 0.1 and significant difference (P = 0.02) between the mean IAP of the first and last min in trial 4. The loss of intra-articular pressure associated with time and motion was 10.5, 16.9, 28.1 and 28.9% for trials 1-4, respectively. As initial intraarticular pressure and motion increased, the percent loss of intra-articular pressure increased.The angle of lowest pressure was 12.2 +/- 1.2 (mean +/- s.e.) in extension in trials 1 and 2, In trials 3 and 4, the lowest pressures were obtained in flexion with the joints at 18.5 +/- 2.0 degrees (mean +/- s.e.). This demonstrated that the joint angle of least pressure changed as the initial intra-articular pressure changed and there would not be a single angle of least pressure for a given joint.The volume of synovial fluid recovered from the MTP joints in trial 3 compared to 4 (trials in which fluid was injected to attain IAP of 30 mmHg) was not significantly different, supporting a soft tissue compliance change as a cause for the significant loss of intra-articular pressure during the 15 min of trial 4.The pressure profiles generated correlate well with in vivo values and demonstrated consistent pressure profiles. Our conclusions are summarised as follows:1. Clinically normal equine MTP joints which were frozen and then later thawed were found to have mostly negative baseline intra-articular pressures, as would be expected in living subjects,2. Alternate pressure profiles of the dorsal and plantar pouch at baseline intra-articular pressure document the presence of pressure forces that would support 'back and forth' fluid movement between joint compartments. This should result in movement of joint fluid during motion, assisting in lubrication and nutrition of articular cartilage,3. If joint pressure was initially greater than normal (30 mmHg), as occurs in diseased equine MTP joints, joint motion further increased joint capsule relaxation (compliance) and, therefore, reduced intra-articular pressure.4. Peak intra-articular pressures reached extremely high values (often >100 mmHg) in flexion when initial pressure was 30 mmHg. Joint effusion pressures recorded for clinical MCP joints are frequently 30 mmHg. These IAP values are expected to produce intermittent synovial ischaemia in clinical cases during joint flexion.5, Additional in vivo studies are necessary to confirm our conclusions from this study and to identify the contributions of fluid absorption and the presence of ischaemia in a vascularised joint.

Magnesium sulphate given topically by iontophoresis for viability of random skin flaps in rats

Our aim was to assess the effects of magnesium sulphate given by iontophoresis on the viability of random skin flaps in rats. Endovenous magnesium sulphate is used to treat pre-eclampsia and diseases of blood vessels. Iontophoresis is an electrotherapeutic method which has shown satisfactory results in controlling ischaemia within the boundaries of the area in which it was given. Forty-five adult male Wistar rats, weighing 300 to 440 g were randomly divided into three groups of 15 animals each: random skin flap (control); random skin flap treated with magnesium sulphate without electrical stimulation; and random skin flap treated with magnesium sulphate with electrical stimulation of 4 mA for 20 minutes. The treatments were applied immediately after the operation and repeated on the following two days. The percentage of necrotic area was measured on the seventh postoperative day using a paper template. For each group, the mean percentage of flap necrosis was as follows: control, 46%; magnesium sulphate without electrical stimulation, 34%; and magnesium sulphate with electrical stimulation, 42%. There was no significant difference among the groups (p=0.18). Magnesium sulphate given by iontophoresis does not increase the viability of random skin flaps in rats.

Comparison of three systems of classification in predicting the outcome of diabetic foot ulcers in a Brazilian population

Objective: The aim was to compare there ulcer classification systems as predictors of the outcome of diabetic foot ulcers; the Wagner, the University of Texas (UT) and the size (area, depth), sepsis, arteriopathy, denervation system (S(AD)SAD) systems in specialist clinic in Brazil.Methods: Ulcer area, depth, appearance, infection and associated ischaemia and neuropathy were recorded in a consecutive series of 94 subjects. A novel score, the S(AD)SAD score, was derived from the sum of individual items of the S(AD)SAD system, and was evaluated. Follow-up was for at least 6 months. The primary outcome measure was the incidence of healing.Results: Mean age was 57.6 years; 57 (60.6%) were made. Forty-eight ulcers (51.1%) healed without surgery; 11 (12.2%) subjects underwent minor amputation. Significant differences in terms of healing were observed for depth (P = 0.002), infection (P = 0.006) and denervation (P = 0.002) using the S(AD)SAD system, for UT grade (P = 0.002) and stage (P = 0.032) and for Wagner grades (P = 0.002). Ulcers with an S(AD)SAD score of <= 9 (total possible 15) were 7.6 times more likely to heal than scores >= 10 (P < 0.001).Conclusions: All three systems predicted ulcer outcome. The S(AD)SAD score of ulcer severity could represent a useful addition to routine clinical practice. The association between outcome and ulcer depth confirms earlier reports. The association with infection was stronger than that reported from the centres in Europe or North America. The very strong association with neuropathy has only previously been observed in Tanzania. Studies designed to compare the outcome in different countries should adopt systems of classification, which are valid for the populations studied.

Efeitos da Cetamina S(+) em dose subanestésica sobre a função e a histologia renal, em modelo de isquemia e reperfusão em ratos

Pós-graduação em Anestesiologia - FMB

The effect of a minor constituent of essential oil from Citrus aurantium: The role of beta-myrcene in preventing peptic ulcer disease

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)