The influence of cardiopulmonary bypass operation on the biodistribution of 99mTc-HMPAO-labelled granulocytes – Evaluation in pigs by planar scintigraphy and section-analyses

Aim of the study was to evaluate the influence of an extra corporal perfusion (cardiopulmonary bypass operation - cpb) on activation and biodistribution of Tc-99m labelled granulocytes in pigs with and without inhibition of the granulocytes by a leukocyte inhibition module (LIM). The cpb is often related to an activation of granulocytes resulting in an inflammatory answer. The biological mechanisms are unsolved yet. First trials of our group showed that LIM may inhibit the activation of neutrophils and therefore antagonize a cpb-caused impairment of cardiac function. This study is the continuation of these experiments with a higher number of animals and the focus on scintigraphic imaging. Animals, material, methods: 39 German landrace pigs were subdivided into three groups: group A (control) median sternotomy without cpb, group B with cpb, group C with LIM in addition to cpb. After labelling with Tc-99m-HMPAO autologues granulocytes were reinjected. Subsequently to cpb, the animals underwent scintigraphic imaging. Quantification was performed with ROI evaluation and with tissue samples (section analysis) examined in a well counter. Results:A high uptake of Tc-99m-HMPAO was found in the liver. The count rates in brain, heart, lung, spleen and kidneys were far below. The amount of Tc-99m-activity in the organ related to the half life corrected administered activity [%] was for the tissue samples (group A/B/C): brain 0.01/0.02/0.03; lung 12.1/8.3/11.5; heart 0.35/0.54/0.42; kidney 1.24/0.87/1.02; spleen 4.0/4.0/4.5, liver 16.8/20.9/19.6. The count rates determined by ROI-evaluation of the scintigraphic images related to the total count rate in the image [%] were (group A/B/C): brain 1.1/0.9/1.0; lung 15.6/10.4/12.2; heart 4.0/3.5/3.4; kidney 4.0/2.9/3.2; spleen 7.6/7.7/9.5, liver 23.1/36.7/31.4. A significant difference in the tracer uptake between the groups could neither be detected by scintigraphic imaging nor evaluation of tissue samples. Conclusion: Scintigraphic imaging as well as section analysis showed a comparable biodistribution of the tracer. Therefore, the initial results of our group were not confirmed with a considerably higher number of animals. Neither cpb nor the use of the LIM influenced distribution of Tc-99m-labelled granulocytes in pigs significantly.

Selective Ablation of the Androgen Receptor in Mouse Sertoli Cells Affects Sertoli Cell Maturation, Barrier Formation and Cytoskeletal Development

The observation that mice with a selective ablation of the androgen receptor (AR) in Sertoli cells (SC) (SCARKO mice) display a complete block in meiosis supports the contention that SC play a pivotal role in the control of germ cell development by androgens. To delineate the physiological and molecular mechanism responsible for this control, we compared tubular development in pubertal SCARKO mice and littermate controls. Particular attention was paid to differences in SC maturation, SC barrier formation and cytoskeletal organization and to the molecular mediators potentially involved. Functional analysis of SC barrier development by hypertonic perfusion and lanthanum permeation techniques and immunohistochemical analysis of junction formation showed that SCARKO mice still attempt to produce a barrier separating basal and adluminal compartment but that barrier formation is delayed and defective. Defective barrier formation was accompanied by disturbances in SC nuclear maturation (immature shape, absence of prominent, tripartite nucleoli) and SC polarization (aberrant positioning of SC nuclei and cytoskeletal elements such as vimentin). Quantitative RT-PCR was used to study the transcript levels of genes potentially related to the described phenomena between day 8 and 35. Differences in the expression of SC genes known to play a role in junction formation could be shown from day 8 for Cldn11, from day 15 for Cldn3 and Espn, from day 20 for Cdh2 and Jam3 and from day 35 for ZO-1. Marked differences were also noted in the transcript levels of several genes that are also related to cell adhesion and cytoskeletal dynamics but that have not yet been studied in SC (Actn3, Ank3, Anxa9, Scin, Emb, Mpzl2). It is concluded that absence of a functional AR in SC impedes the remodeling of testicular tubules expected at the onset of spermatogenesis and interferes with the creation of the specific environment needed for germ cell development.

Cinética do transporte jejunal de glicose em ratos com deficiência de niacina.

The kinetic of jejunal glucose transport was studied using perfused rat jejunum in vivo. Ninety rats were fed a diet deficient in niacin and 90 a control diet. The jejunal loops of 7 groups of animal were infused each group with one of following solutions of glucose: 5, 10, 20, 40, 80, 160 and 300 mM/l. The Vmax and Km values were determined. The results showed that the vitamin-deficient rats absorbed less glucose independently of the amount infused and these animals had lower Vmax (133.7 microM/15 min/15 cm) and Km (192.1 mM/l) than control groups (294.1 microM/15 min/15 cm and 171.8 mM/l, respectively). In conclusion one can assume that niacin deficiency leads to a decreased glucose absorption in the jejunal loops, when tested as in our experimental model.

A influência da perfusão cardíaca nas relações pressão/volume do ventrículo esquerdo.

PURPOSE--To analyze the influence of the myocardial engorgement due to coronary perfusion on the left ventricular diastolic pressure/volume (P/V) relations. METHODS--The study was undertaken in the isovolumic blood-perfused dog heart preparation (n = 7). The P/V relations were determined promoting ventricular volumes variations by steps of 2ml, in two conditions: during coronary perfusion pressure of 100mmHg and after occlusion of the perfusion line. RESULTS--It was verified that the P/V relations obtained without myocardial perfusion shifted to the down right in respect to the perfused heart. A exponential fitting of the P/V relations allows to conclude that there was no difference among the slopes of the curves obtained with and without perfusion. CONCLUSION--Our results indicated that coronary perfusion restrain the capacity of the left ventricle to receive blood, but there is no influence on the passive elastic stiffness of the chamber. This effect was taught to be consequence of the myocardial erectile property.

INFLUENCIA DO ACRESCIMO DE MANITOL A SOLUCAO NUTRIENTE NO DESEMPENHO MECANICO E NO GRAU DE EDEMA MIOCARDICO DE CORACOES ISOLADOS DE RATOS

Purpose - To analyse the influence of mannitol added to Krebs-Henseleit (KH) solution on the myocardium edema and myocardial function. Methods - Isolated rat heart under isovolumetric contractions studied according to Langendorff's technique were perfused with KH solution at constant flow during 90 min. The coronary perfusion pressure, diastolic and systolic pressures were recorded at every 15 min. At the end of the experiment, myocardium water content was measured in hearts perfused with KH solution (group I, n = 9) and in hearts perfused with KH solution plus 8 mM mannitol (group II, n = 8). These results were compared to non-perfused control heart (n = 9). Results - Myocardial water content was statistically higher in group I (80.8 ± 1.3%) compared to group II (78.1 ± 0.7%) and control group (75.5 ± 0.5%). Systolic arterial pressure was statistically higher in group I (86.2 ± 11.5 mmHg) compared to group II (72.7 ± 21.1 mmHg). There was no difference in the diastolic pressure between the two groups. Coronary perfusion pressure (Pp) increased progressively during the experiment in both groups. However, Pp was lower in group II than in group I. Conclusion - Mannitol added to KH solution significantly attenuates the myocardium edema in the isolated perfused rat heart.

Influencia da elevacao sustentada da pressao arterial sobre a dP/dt do ventriculo esquerdo mantendo-se constante a pressao diastolica do ventriculo esquerdo

Purpose - To evaluate the influence of sustained elevations of arterial pressure on dP/dt values, which the left ventricular end diastolic pressure was kept constant. Methods - Thirteen anesthetized dogs, mechanically ventilated and submitted to thoracotomy and pharmacological autonomic block (atropine - 0.5 mg/kg IV + oxprenolol - 3 mg/kg IV) were studied. The arterial pressure elevation was obtained by mechanical constriction of the descending thoracic aorta. Analyses were made in control (C) situation and after two successives increments of arterial pressure, sustained for 10min, called hypertension 1 (H1) and hypertension 2 (H2), respectively. The end diastolic left ventricular pressure was kept constant by utilization of a perfusion system connected to the left atria. Results - Heart rate did not change (C: 125 ± 13.9bpm; H1: 125 ± 13.5bpm; H2: 123 ± 14.1bpm; p > 0.05); the LVSP increased (C: 119 ± 8.1mmHg; H1: 142 ± 7.9mmHg; H2: 166 ± 7.7mmHg; p < 0.01); the AoDP increased (C: 89 ± 11.6mmHg; H1: 99 ± 9.5mmHg; H2: 120 ± 11.8mmHg; p < 0.01); the LVEDP (C: 6.2 ± 2.48mmHg; H1: 6.3 ± 2.43mmHg; H2: 6.1 ± 2.51mmHg; p > 0.05) and the dP/dt (C: 3068 ± 1057.1mmHg/s; 3112 ± 995.7mmHg/s; H2: 3086 ± 979.5mmHg/s; p > 0.05) did not change. Conclusion - dP/dt values are not influenced by a sustained elevation of arterial pressure, when the end diastolic left ventricular pressure is kept constant.

Effect of α-tocopherol, taurine and selenium on the attenuation of ischemia/reperfusion injury of splanchnic organs

Background: Splanchnic artery occlusion shock is caused by increased capillary permeability and cellular injury precipitated by oxygen derived free radicals following ischemia and reperfusion of splanchnic organs. The purpose of this study was to assess the role of several well-known oxygen- derived free radical scavengers in ameliorating or preventing this syndrome. Study design: Anesthetized rats were subjected to periods of occlusion of the visceral arteries and reperfusion. Tocopherol, taurine, selenium or a 'cocktail' of these three agents was injected subcutaneously for 4 consecutive days prior to operation. Mean arterial blood pressure was measured throughout the experimental period. Fluorometry and technetium-99m pyrophosphate counting of the visceral organs were performed as well as a histologic grading system for intestinal viability. Results: Final mean arterial blood pressure associated with the 'cocktail' and selenium groups was 79.1 ± 27.4 mmHg and 83.6 ± 17.8 mmHg, respectively. These values were significantly higher than the control group, 40.8 ± 11.4 mmHg (P < 0.05). Similar patterns of the benefit of selenium in contrast with the other groups were obtained with fluorescein perfusion, radioisotopic activity and histologic analysis. Conclusion: Pretreatment with selenium of splanchnic ischemia and reperfusion in the rat improves mean arterial blood pressure and microcirculatory visceral perfusion. Further analysis of the precise protective mechanism of selenium for reperfusion injury will enable visceral organs to withstand the consequences of increased capillary leakage and oxidant injury.

A lung computed tomographic assessment of positive end-expiratory pressure-induced lung overdistension

The aim of this study was to assess positive end-expiratory pressure (PEEP)-induced lung overdistension and alveolar recruitment in six patients with acute lung injury (ALI) using a computed tomographic (CT) scan method. Lung overdistension was first determined in six healthy volunteers in whom CT sections were obtained at FRC and at TLC with a positive airway pressure of 30 cm H2O. In patients, lung volumes were quantified by the analysis of the frequency distribution of CT numbers on the entire lung at zero end-expiratory pressure (ZEEP) and PEEP. In healthy volunteers at FRC, the distribution of the density histograms was monophasic with a peak at -791 ± 12 Hounsfield units (HU). The lowest CT number observed was -912 HU. At TLC, lung volume increased by 79 ± 35% and the peak CT number decreased to -886 ± 26 HU. More than 70% of the increase in lung volume was located below -900 HU, suggesting that this value can be considered as the threshold separating normal aeration from overdistension. In patients with ALI, at ZEEP the distribution of density histograms was either monophasic (n = 3) or biphasic (n = 3). The mean CT number was -319 ± 34 HU. At PEEP 13 ± 3 cm H2O, lung volume increased by 47 ± 19% whereas mean CT number decreased to -538 ± 171 HU. PEEP induced a mean alveolar recruitment of 320 ± 160 ml and a mean lung overdistension of 238 ± 320 ml. In conclusion, overdistended lung parenchyma of healthy volunteers is characterized by a CT number below -900 HU. This threshold can be used in patients with ALI for differentiating PEEP-induced alveolar recruitment from lung overdistension.

Distúrbios de memória em pacientes epilépticos

The author studied the mnemonic activity from epileptic patients with complex partial seizures (CPS), with the aims: 1) to identify memory disorders; and 2) to compare the patients' with the controls' results. Fifty adult patients and 20 subjects without neuropsychiatric disorders were studied. The methods consisted in: 1) investigation of the mnemonic activity through the Wechsler Memory Test (subtests: Storage and Recall, Recent Memory, and Immediate Memory); 2) comparison among the results of both groups; association from mnemonic activity with brain SPECT. In the three subtests, the patients showed cognitive performance significantly lower than the controls (p <0,05). It was found association from reduced blood flow, mainly in left temporal region, with memory impairment of the three subtetsts. The conclusion was that the CPS are associated to memory impairment.

Efeitos do alopurinol em rins isquemicos de caes anestesiados com pentobarbital sodico

Background and Objectives - Allopurinol is a drug which inhibits the formation of noxious renal free radicals. The aim of this study was to evaluate the protecting renal effects of allopurinol in ischemic kidneys of dogs. Methods - Sixteen dogs were anesthetized with sodium pentobarbital and submitted to extracellular volume expansion (1.4 ml.kg-1.min-1), to mechanic ventilation with air, to right nephrectomy and to left renal artery clamping. Changes which might occur in renal morphology and function after 30 min of total ischemia and posterior reperfusion were studied in Group 1 (G1), in addition to the action of allopurinol (50 mg.kg-1) on those kidneys, when administered 24 h before the experiment and 1 h before the ischemic procedure in Group 2 (G2). The following parameters: heart rate, inferior vena cava pressure, mean blood pressure, PAH clearance (PAH(c)), renal blood flow (RBF), renal vascular resistance (RVR), creatinine clearance (Cr(c)), filtration fraction, urine output, plasma and urine osmolality, osmolar clearance, free water, sodium and potassium clearance, urine and fractional sodium and potassium excretion, hematocrit, rectal temperature, and left kidney histology were evaluated in four moments: M1 control, and M2, M3, M4 obtained immediately, 15 and 30 min after unclamping of the left renal artery. In G2, M1, M2, M3 and M4 were obtained 45, 90, 105, and 120 min after the second allopurinol dose. Results - Both groups showed the highest values for PAH(c), RBF, and Cr(c), and the lowest values for RVR in M1. Animals were tachycardiac since the beginning of the experiment both in G1 and in G2. The other parameters were not changed. Left kidney histological evaluation showed alterations compatible with acute tubular necrosis in both experimental groups. Conclusions - Alterations found in renal hemodynamics were compatible with the release of vasoconstrictor substances due to renal ischemia. Allopurinol was not effective in preventing renal alterations caused by ischemia and reperfusion.

Scanning electron microscopy study of the choroid plexus in the monkey (Cebus apella apella)

The cells of the choroid plexus of the lateral ventricles of the monkey Cebus apella apella were examined through scanning electron microscopy at contributing to the description of such structures in primates. The animals were anesthetized previously with 3% hypnol intraperitoneally and after perfusion with 2.5% glutaraldehyde, samples of the choroid plexus were collected after exhibition of the central portion and inferior horn of the lateral ventricles. The ventricular surface of those cells presents globose form as well as fine interlaced protrusions named microvilli. Among those, it is observed the presence of some cilia. Resting on the choroid epithelial cells there is a variable number of free cells, with fine prolongations which extend from them. They are probably macrophages and have been compared to Kolmer cells or epiplexus cells, located on choroid epithelium. The choroid plexus of the encephalic lateral ventricles of the monkey Cebus apella apella at scanning electron microscopy is similar to that of other primates, as well as to that of other species of mammals mainly cats and rats, and also humans.

Efeito de drogas utilizadas no tratamento de hipertensão arterial sistêmica sobre a pressão intra-ocular: Estudo experimental no cão

Purpose: To study the effects of two drugs (captopril and propranolol) used in the treatment of systemic hypertension, on the intraocular pressure (IOP) of anesthetized dogs. Methods: 24 dogs, divided into 3 groups of 8 each. In the first group, 1.5 mg/kg IV of captopril (an angiotensin converting enzyme inhibitor) was administered. In the second group, 1.5 mg/kg IV of propranolol (a beta-blocker) was administered. The third group was the control. IOP and blood pressure (BP) were measured by manometry. The perfusion pressure was calculated by the difference between BP and IOP (BP-IOP). The parameters were studied at 6 moments (0, 10, 30, 60, 90 and 120 minutes). Results: There was significant reduction of IOP (p<0.05) with captopril and propranolol, without difference between the drugs. With captopril the BP and PP decreased markedly at 10 and 30 minutes. With propranolol there was no reduction of BP or PP. Conclusions: Captopril and propranolol reduced IOP. However, the marked reduction of BP, and consequently of PP caused by captopril may be undesirable for irrigation of the optic nerve.

Efeitos do extrato aquoso de cebola (Allium cepa L.) sobre a função renal e a pressão arterial em ratos Wistar

The vegetal species, Allium cepa, known as onion, is widely used in the folk medicine as diuretic, besides it has been used on the bronchitis, cough, cardiovascular diseases and hypertension treatment. In this study we evaluate the onion aqueous extract (AE) effect on water flow and electrolytes in anesthetized Wistar rats, besides we also evaluate arterial pressure alterations. Two groups were studied: Group 1 (control) - oral tratment with 1.0 mL of distilled water, and Group 2 (experimental) - oral treatment with 1.0 mL of AE 20%. The rats were anesthetized and we canulate the trachea, left carotide artery (for arterial pressure measurement and blood collecting), jugular vein (to execute inulin perfusion - to register glomerular filtration), and urinary bladder (to collect urine). The Group 1 results had shown that the animals had not presented significant alterations (p>0.05) in the analyzed parameters. The animals of Group 2 had a significant reduction (p<0.05) in the arterial pressure (22.0%). However, there were not significant alterations in renal parameters (p>0.05). These results show that the treatment with the AE lead a hypotensor effect in anesthetized Wistar rats, but not followed by renal parameters alterations.

Action of anti-bothropic factor isolated from Didelphis marsupialis on renal effects of Bothrops erythromelas venom

Acute renal failure is the most common complication in the lethal cases caused by snakebites in Brazil. Among the Brazilian venom snakes, Bothrops erythromelas is responsible for the majority of accidents in Northeastern Brazil. Didelphis marsupialis serum could inhibit myonecrotic, hemorrhagic, edematogenic hyperalgesic and lethal effects of envenomation determined by ophidian bites. In the present study, we evaluated the action of the anti-bothropic factor isolated from D. marsupialis on the renal effects promoted by B. erythromelas venom without systemic interference. Isolated kidneys from Wistar rats were perfused with Krebs-Henseleit solution containing 6% bovine serum albumin. We analyzed renal perfusion pressure (PP), renal vascular resistance (RVR), glomerular filtration rate (GFR), urinary flow (UF), and the percentages of sodium and potassium tubular transport (%TNa +, %TK +). The B. erythromelas venom (10 μg mL -1) decreased the PP (ct=108.71±5.09 mmHg; BE=65.21±5.6 mmHg*) and RVR (ct=5.76±0.65 mmHg mL -1 g -1 min -1; BE=3.10±0.45 mmHg mL -1 g -1 min -1*) . On the other hand, the GFR decreased at 60 min (ct 60=0.76±0. 07 mL g -1 min -1; BE 60=0.42±0.12 mL g -1 min -1*) and increased at 120 min (ct 120=0.72±0.01 mL g -1 min -1; BE 120=1.24±0.26 mL g -1 min -1*). The UF increased significantly when compared with the control group (ct=0.14±0.01 mL g -1 min -1; BE=0.47±0.08 mL g -1 min -1*). The venom reduced the %TNa + (ct 90=79.18±0.88%; BE 90=58.35±4.86%*) and %TK + (ct 90=67.20±4.04%; BE 90=57. 32±5.26%*) The anti-bothropic factor from D. marsupialis (10 μg mL -1) incubated with B. erythromelas venom (10 μg mL -1) blocked the effects on PP, RVR, %TNa +, and %TK +, but was not able to reverse the effects in UF and GFR promoted by venom alone. However, the highest concentration of D. marsupialis serum (30 μg mL -1) reversed all the renal effects induced by the venom. In conclusion, B. erythromelas venom altered all the renal functional parameters evaluated and the anti-bothropic factor from D. marsupialis was able to inhibit the effects induced by the venom in isolated kidney. © 2005 Elsevier Ltd. All rights reserved.