Quantifying Single Microvessel Permeability in Isolated Blood-perfused Rat Lung Preparation

The isolated blood-perfused lung preparation is widely used to visualize and define signaling in single microvessels. By coupling this preparation with real time imaging, it becomes feasible to determine permeability changes in individual pulmonary microvessels. Herein we describe steps to isolate rat lungs and perfuse them with autologous blood. Then, we outline steps to infuse fluorophores or agents via a microcatheter into a small lung region. Using these procedures described, we determined permeability increases in rat lung microvessels in response to infusions of bacterial lipopolysaccharide. The data revealed that lipopolysaccharide increased fluid leak across both venular and capillary microvessel segments. Thus, this method makes it possible to compare permeability responses among vascular segments and thus, define any heterogeneity in the response. While commonly used methods to define lung permeability require postprocessing of lung tissue samples, the use of real time imaging obviates this requirement as evident from the present method. Thus, the isolated lung preparation combined with real time imaging offers several advantages over traditional methods to determine lung microvascular permeability, yet is a straightforward method to develop and implement.     

High-dose mode of death in Tribolium

This study reports the first demonstration within a single insect genus (Tribolium) of both the acute, or lethal-midlethal, dose-independent pattern of mortality, and the hyperacute, dose-dependent pattern, after appropriate doses of ionizing radiation. This demonstration provides resolution of apparently contradictory reports of insect responses in terms of doses required to cause lethality and those based on survival time as a function of dose. A dose-dependent mortality pattern was elicited in adult Tribolium receiving high doses, viz., 300 Gy or greater; its time-course was complete in 10 days, before the dose-independent mortality began. Visual observations of heavily-irradiated Tribolium suggested neural and/or neuromuscular damage, as had been previously proposed by others for lethally-irradiated wasps, flies, and mosquitoes. Results of experiments using fractionated high doses supported the suggestion that the hyperacute or high-dose mode of death is the result of damage to nonproliferative tissues. Relative resistance of a strain to the hyperacute or high-dose mode of death is not necessarily correlated with resistance to the midlethal mode, which is believed to be the result of damage to the proliferative cells of the midgut.

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Résumé Les résultats de nombreuses études des réactions des insectes adultes de différents groupes à l'irradiation s'opposent quant à l'importance de la dose provoquant la létalité et quant aux modalités de la mort. Les diptères et les guèpes impliquent des doses très élevées,-des centaines de Gy-, ne présentent aucune période caractérisant la mort par irradiation, et décèdent de plus en plus tôt avec l'augmentation des doses. Beaucoup d'autres insectes succombent à des doses (milétalelétale) beaucoup plus faibles,-de quelques Gy à des dizaines-, et quelle que soit la dose meurent au bout d'un temps voisin.Au cours de cette étude, nous avons pu observer que ces deux types de mortalité peuvent être provoqués chez le même genre d'insecte (Tribolium), avec des doses convenables d'irradiation . Un syndrôme caractéristique a été provoqué avec des doses très élevées, de 300 Gy ou plus,-à ces doses la mort est obtenue en 10 jours après l'irradiation. L'absence de syndrôme caractéristique se produit avec des doses inférieures ou égales à 80 Gy; la mort a lieu alors entre 10 et 16 jours en fonction de la dose.Les différences entre les deux types de décès indiquent deux processus de mort par irradiation. La manifestation d'une désorientation et d'une perte de coordination motrice chez les Tribolium fortement irradiés suggère des altérations neurales et/ou neuromusculaires comme cause/s de ce type de mort provoquée par des doses élevées. L'implication de tissus sans prolifération a été confortée par les résultats d'expériences utilisant de hautes doses fractionnées. Le type de mort milétal est considéré après de nombreuses observations indépendantes, comme le résultat d'atteintes à la prolifération des cellules de l'intestin moyen.Les données contradictoires sur les réactions des insectes aux irradiations proviennent d'abord de l'absence de connaissances sur le lieu des dégâts. Les diptères sont connus maintenant, après différentes études, comme perdant complètement l'aptitude au renouvellement cellulaire, et présentent ainsi un type de mort avec dose élevée. Beaucoup d'autres insectes adultes ont un renouvellement cellulaire de l'intestin moyen limité, et ainsi présentent le type de mort milétal. Le type de mort, dit haute dose, peut être induit dans cette dernière catégorie d'insectes par une irradiation suffisamment forte, et, dans le cas du Tribolium le déroulement de la mort se produit alors de deux façons bien distinctes.

     

Disturbed lipid metabolism in diabetic coronary vessels

The aim of this study was to clarify whether or not arachidonic acid metabolic disorders are caused by a substrate inavailability and whether such disorders might contribute to circulatory disturbances in the diabetic myocardium. Norepinephrine induced a decrease in the conductivity of both coronary arterial bed and myocardial microcirculation in alloxan-diabetic dogs. It was markedly (p < 0.05) attenuated both by indomethacin and acetylsalicylic acid pretreatments indicating an imbalance among the vasoactive prostanoids in diabetes. TXA₂ release from the diabetic coronary rings was found to be elevated and could be normalized after the blockade of vascular adrenoceptors by phentolamine (p < 0.05). PGIZ synthesis was also enhanced by adrenergic blockade in the diabetic arterial rings. After pretreatment with ^l4C arachidonic acid, in order to measure substrate availability, the arachidonic acid metabolic rate was less in the diabetic coronary arteries than in healthy vessels (p < 0.05). Ten µmol/1 norepinephrine decreased arachidonic acid metabolism in the presence of prelabelled substrate in the diabetic animals, compared to an increase observed in metabolically healthy dogs. Therefore diabetes appears to diminish arachidonic acid metabolism and uptake independent of adrenoceptors and to induce an imbalance between vasoconstrictor and vasodilator cyclooxygenase products, resulting in elevated TXA₂ release controlled by adrenergic mechanisms which may contribute to an impairment in myocardial microcirculation.Abbreviations 6-oxo-PGF₁ 6-oxo prostaglandin F₁ - HPLC High Pressure Liquid Chromatograph - LAD Left Anterior Descending (coronary artery) - PGI₂ Prostacyclin - TXA₂ Thromboxane     

Carbohydrates,lipids and lipoproteins and islet changes in diabetes with superimposed myocardial infarction

Short-term metabolic and concomitant morphologic effects of streptozotocin diabetes on isoproterenol-induced myocardial infarction was studied in Wistar rats, Of particular significance was the observation that myocardial infarction in concert with diabetes brought about a distinctive exacerbation of the severity and complexity of the histopathological lesions. Of all the biochemical parameters, serum glucose and free fatty acids registered maximum elevation and serum lactate and cardiac glycogen levels a maximum reduction. Among the lipoproteins, an inverse relationship was found between high density lipoproteins and low density and very low density lipoproteins; while high density lipoproteins, ratio of high density lipoprotein to low density lipoprotein and the percentage of high density lipoprotein were decreased, there was a significant increase in low density lipoprotein concentration and percentage values of low density and very low density lipoproteins. In diabetes, the B cell of the endocrine pancreas depicted selective necrosis. Loss of insulin granules and wide-spread necrobiosis of cellular elements of the pancreatic islets were observed, respectively, in myocardial infarction and in diabetes plus myocardial infarction combinations. Pathological evidence of chemical-induced mild toxicity was present in the exocrine parenchyma. Mitotic features and the presence of centroacinar cells in the damaged Langerhans’ islets supposedly formed the basis of regeneration of the tissue in diabetes, with or without vascular complications     

Meteorological factors and the time of onset of chest pain in acute myocardial infarction

Analysis of the time of onset of chest pain in 2254 patients with a myocardial infarction admitted to a coronary care unit in Leicester during a 10-year period shows an association with temperature and humidity. During both the most cold and humid times of the year, the relationship is a strong one. A generalized linear model with a log link was used to fit the data and the backward elimination selection procedure suggested a humid, cold day might help to trigger the occurrence of myocardial infarction. In addition, cold weather was found to have a stronger effect on the male population while those men aged between 50 and 70 years were more sensitive to the effect of high humidity.     

急性低氧对大鼠血液中儿茶酚胺及血小板聚集性的影响

健康ＳＤ雄性大鼠,体重２５０－３００ｇ,麻醉、气管插管,用人工呼吸机经气袋供气,自发吸入氧浓度为９％的氧氮混合气,用高效液相色谱－电化学联合检测法及电阻法检测循环血液中儿茶酚胺及全血血小权聚集性的动态变化。结果：急性低氧１５ｍｉｎ时血液中肾肾上腺素（Ａ）浓度及全血血小板聚集性显著增加（Ｐ〈０．０１）,而去甲肾上腺素（ＮＡ）浓度虽有所增加,但无统计学意义（Ｐ〉０．０５）;复氧１５ｍｉｎ时血液中儿茶酚     

Cardiac phosphocreatine deficiency induced by GPA during postnatal development in rat

The effect of chronic administration of -guanidinopropionic acid (GPA) on the protein profiling, energy metabolism and right ventricular (RV) function was studied in the rat heart during the weaning and adolescence period. GPA was given in tap water (1–1.5%) using pair drink controls. The feeding of animals with GPA solution for a six week period resulted in elevation of heart to body weight ratio due to body growth retardation. GPA accumulated in the myocardium up to 67.37 ± 5.3 moles.g dry weight and the tissue content of total creatine, phosphocreatine and ATP was significantly decreased to 15%, 9% and 65% of control values respectively. Total activity of creatine kinase (CK) was not changed, but the proportion of mitochondrial (Mi) CK isoenzyme was decreased; the percentage of MB isoenzyme of CK was significantly higher. GPA treatment resulted in an elevation of the content of cardiac collagenous proteins and decrease of non-collagenous proteins in the heart; in parallel, a decrease of the collagen I to collagen III ratio was detected. The function of the RV was assessed using an isolated perfused heart with RV performing pressure-volume work. As compared to pair-drink controls, RV function was significantly impaired the GPA group: at any given right atrial filling pressure, the RV systolic pressure and the rate of pressure development were decreased by almost a factor of two. Elevation of the RV diastolic pressure with increasing pulmonary artery diastolic pressure was also significantly steeper in the GPA group which also showed decrease of cardiac output, especially at high outflow resistance. It may be assumed that chronic administration of GPA deeply influenced metabolic parameters, protein profiles and contractile function of the developing heart. On the other hand, concentrations of glucose, total lipids and triglycerides in blood plasma were not affected. All these data confirm the concept that the CK system is of central importance both for heart function and for the regulation of normal growth of cardiac myocytes.     

The ‘World Prodigy’ oil spill in Narragansett Bay,Rhode Island: acute effects on macrobenthic crustacean populations

On June 26,1989, the tanker World Prodigy ran aground just outside the mouth of the West Passage of Narragansett Bay, Rhode Island, USA. About 922 metric tons of No. 2 fuel oil were released into the water and drifted over a total area of about 120 sq miles. Three days after the spill only a small fraction of the oil remained.The effects on macrobenthic crustaceans within the first five weeks after the spill were studied at five stations with a varying degree of oil exposure, including one control site never reached by oil from the spill. Significant differencies between these stations were noted for total amphipod abundance, the amphipod genus Ampelisca and ostracods (retained on a 0.3 mm mesh), but not for amphipods of the genus Corophium. At the most heavily impacted station (23 µg oil g^–1 sediment dry weight), the total amphipod abundance, dominated by Ampelisca verrilli, decreased by 86% within the first two weeks after the spill. Decreases in total amphipod abundance significantly larger than at the control site were noted also at two other stations, one of which with only trace amounts of oil detected in the sediment. The amphipod populations at these sites were dominated by juvenile specimens.These findings confirm the extreme sensitivity to oil pollution of amphipods and ostracods, noted in earlier field and experimental studies.