Immunohistochemical study of myoglobin deletion from myocardium in hemorrhagic shock

Myoglobin (MB) deletion from myocardium in the case of hemorrhagic shock was firstly studied by immunohistochemical and morphometry technique. The results showed that there were different degrees of segmental deletion of MB from myocardium in each case of hemorrhagic shock which has continued over an hour. The significance of these changes in forensic medicine are discussed.     

Reevaluation of the linkage between acute hemorrhagic shock and bacterial translocation in the rat

The present study was undertaken to determine the conditions under which acute periods of hemorrhagic shock induce bacterial translocation. Rats (at least six per group) were anesthetized intraperitoneally with the barbiturate, pentobarbital (50 or 65 mg/kg), or the inhalation anesthetic methoxyflurane. Following anesthesia, the femoral artery was catheterized, from which blood was withdrawn to maintain a mean arterial blood pressure of 30 mmHg for 30, 60, or 90 min, followed by reinfusion of shed blood. Instrumented, but nonshocked animals served as controls. Rats were sacrificed at 0, 2, or 24 hr postshock, and quantitative bacterial cultures of the mesenteric lymph node complex (MLN), liver, and spleen were made. Within groups, the effects of heparinization were also determined. In pentobarbital-treated animals, regardless of the extent of heparinization, consistent translocation to both MLN and distant organs occurred when shock was prolonged for 90 min, and assessment of translocation was made 24 hr after reinfusion of shed blood. Furthermore, a mortality rate of approximately 30% was found in rats subjected to this protocol. The magnitude of translocation was less consistent, and did not differ from that in sham shock controls, under other conditions of shock and evaluation. In rats anesthetized with methoxyflurane, no mortality occurred, and no statistical significance between the incidence or degree of translocation in shocked animals vs. sham shock controls could be demonstrated, regardless of the shock protocol. In additional studies, effects of these anesthetics on intestinal morphology and superior mesenteric arterial (SMA) flow in the context of hemorrhagic shock were assessed.(ABSTRACT TRUNCATED AT 250 WORDS)     

Capsaicin-resistant vagal afferent fibers in the rat gastrointestinal tract: anatomical identification and functional integrity

The presence and distribution of vagal fibers and terminals throughout esophagus and gastrointestinal tract that could be anterogradely labeled by nodose ganglion tracer injections was quantitatively assessed in capsaicin- and vehicle-pretreated adult rats, in order to identify the capsaicin-resistant population. Up to 90% of the intraganglionic laminar endings (IGLEs), in the myenteric plexus of the esophagus, and 70-90% in the stomach, as well as 57% of the intramuscular endings or arrays (IMAs) in the fundic stomach survived the capsaicin treatment, while in the upper small intestine only few and in the lower small intestine, the cecum and colon, virtually no IGLEs survived capsaicin treatment. Intramucosal terminals were not assessed. Furthermore, gastric balloon distension-induced c-Fos expression in the dorsal vagal complex was not significantly decreased in capsaicin-treated rats. It is concluded that among primary vagal afferents there is a capsaicin-resistant population that primarily innervates the esophagus and upper gastrointestinal tract, and a capsaicin-sensitive population that innervates mainly the lower tract. At least vagal gastric tension-sensitive afferents also seems to be functionally intact in that they may be capable of synaptically activating second-order neurons in the brainstem.     

Effects of dexamethasone on functional and pathological changes in rat bronchi caused by high acute exposure to chlorine

OBJECTIVE: To assess month-to-month variability of total cholesterol, triglycerides, high-density lipoprotein-cholesterol (HDL-C), calculated low-density lipoprotein-cholesterol (LDL-C), apolipoprotein A1, apolipoprotein B, and lipoprotein (a), as well as factors that could influence variability, including recent acute infection in an adolescent population. METHODS: Sixty-three high school students had fasting lipids and lipoproteins measured at 4 separate times during the school year and another venipuncture 3 to 7 days after recovery from an acute infection. Erythrocyte sedimentation rate was also measured. Coefficients of variation were calculated for each study variable. The influence of recent infection on variability was assessed. RESULTS: The 50th and 95th percentiles, respectively, for the coefficient of variation for each variable were as follows: total cholesterol, 7.3% and 13.6%; triglycerides, 22% and 47.3%; HDL-C, 7.9% and 16.8%; LDL-C, 12.1% and 25%; apolipoprotein A1, 6.3% and 15.2%; apolipoprotein B, 9.5% and 17.2%; and lipoprotein (a), 19.3% and 40%. Recent infection significantly lowered HDL-C (4 mg/dL; P < .0001) and apolipoprotein A1 (7 mg/dL; P < .005). CONCLUSIONS: Clinicians evaluating lipids and lipoproteins serially should expect significant visit-to-visit variation in triglycerides and calculated LDL-C values. Assessment of HDL-C and apolipoprotein A1 should not be done within 2 weeks of an acute infection. Apolipoproteins B and A1 have slightly less variability than their respective lipoprotein cholesterol values (LDL-C and HDL-C).     

Regulation of cysteine-rich intestinal protein by dexamethasone in the neonatal rat

The cysteine-rich intestinal protein (CRIP) is an intestinal zinc-binding protein containing a single copy of a cysteine-rich domain known as the LIM motif. CRIP mRNA and protein levels increased in the rat small intestine throughout the suckling period, reaching highest levels by the late weanling stage. A similar developmental pattern of CRIP protein levels was also detected by an increase in zinc binding to CRIP-containing HPLC fractions of intestinal cytosol. Administration of the synthetic glucocorticoid hormone dexamethasone to neonates caused the precocious rise of CRIP mRNA and protein. In adult rats, CRIP mRNA levels were not significantly altered by dexamethasone. Maximal CRIP mRNA content was detected in cells from the mid-villus, as confirmed by expression of cryptdin mRNA. In this report we show the glucocorticoid regulation of the LIM motif-containing protein CRIP and suggest that glucocorticoid hormones play a role in developmental regulation of CRIP.     

Potential role of the peroxynitrate-poly(ADP-ribose) synthetase pathway in a rat model of severe hemorrhagic shock

PURPOSE: The purpose of this study was to assess the performance of sealants placed by senior dental students as part of a comprehensive dental care program that included periodic patient recall. METHODS: The dental records of 100 patients ranging in age from 6 to 13 years were selected for review to determine the treatments provided for first permanent molars over time. Criteria for inclusion were: 1) at least five documented recall examinations and 2) all four first permanent molars had to have been treated with an occlusal pit and fissure sealant. The data collected included: 1) the age of the patient at the time of initial sealant placement; 2) the subsequent treatment provided to the first permanent molars, including retreatment with sealant or restoration and the date the services were provided; 3) the last date of follow-up examination in the pediatric dental program. RESULTS: A total of 400 molars were followed for an average of 54 months. Fifty-two percent of all molars received no further treatment after initial placement of sealant. Approximately 35% received retreatment with sealant only. The total number of molars receiving sealant material only was 343 (86%). The total number of teeth that were judged to require restoration was 57 (14%). No relationship was noted between the patient's age at placement of the occlusal sealant and sealant performance. CONCLUSION: In a dental school clinic, occlusal sealants were effective at preventing caries in a comprehensive care program that included periodic recall.     

Function of the liver reticuloendothelial system in hemorrhagic shock in dogs

The function of hepatic reticulo-endothelial system at normal circulation and during hemorrhagic shock was examined in 13 dogs. The results were as follows:1.198Au-colloid liver scintigraphy demonstrated no essential alterations by shock. A trifling diminution caused by the diminished blood amount in the liver was found at heavy blood loss. The spleen was opcified in no case. 2. The elimination rate, resp. the blood clearance, of 198Au-colloid remained constant during the first 2 hours, proving that the vital clearance function of hepatic reticulo-endothelial system is completely maintained in this phase of hemorrhagic shock. After longer duration of hypovolemic shock (5 hours) elimination rate decreased. 3. It can be concluded that hepatic circulation at first alters according to the blood volume. 4. Consequently, functional testing of hepatic reticulo-endothelial system is not fit for early diagnosis of shock.     

Flowmeter changes induced by glucagon in hepato-portal hemodynamics in the course of hemorrhagic shock in dogs

An easily applicable score to predict the risk of prematurity (Tab. I) (defined by weight) is examined prospectively (scoring during 6 th month of pregnancy) in 431 and retrospectively (obtained after delivery) in 1183 pregnancies. In the prospective study (Tab. II) 71.4% of all pregnancies resulting in babies below 2501 g exceed the proposed 50 points risk probabilty limit whereas only 18.7% of pregnancies with babies of more than 3000 g do so. Excluding pregnancies with 20 or more risk points and excellent prenatal care (8 or more consultations) - which should change the outcome of risk-pregnancies - the percentages are 77.8% and 12.2% respectively (Tab. III). Pregnancies resulting in babies with birth weight of 2501 g -2750 g exceeded the limit in 38.9% and those with babies of 2751 g-3000 g in 20.7%. If 60 risk points are used as the limit the percentages for more than 3000 g until less than 2501 g would be 8.2%, 6.9%, 33.3% and 66.7%. In the resrospective study (Tab. V) 14.7% of all pregnancies with babies above 3000 g exceeded the 50 risk points limit compared with 57.2 of those with babies below 2501 g. Excluding pregnancies with 20 or more risk points and excellent prental care the percentages are 7.6 and 59.4 respectively. In the retrospective study the influence of the quality of prenatal care by the number of consultations (3-4; 5-7; 8 or more) is clearly demonstrable: Pregnancies with more than 50 risk points resulted in 80.7%, 57.1% and 19.8% depending on the quality of care in babies below 2501 g. Pregnancies with 31-50 risk points did so in 47.2%, 20.4% and 11.8%. In 334 women the score could be applied twice, in the 6th month and at delivery. Comparing both scores it was found that only 1.8% of these women exceeded the 50 risk points limit by events occurring after the 6th month scoring (Tab. IV). The score, simple enough to be applied by nurses and midwives, seems to be able to select 77.8% of pregnancies resulting in babies below 2501 g already during the 6th month of pregnancy, i.e. early enough for preventive measures to be taken that decrease the frequency of underweight births by three quarters.     

Alveolar liquid clearance is increased by endogenous catecholamines in hemorrhagic shock in rats

The primary objective of this study was to test the hypothesis that hemorrhagic shock would stimulate alveolar liquid clearance by a catecholamine-dependent mechanism. Anesthetized rats were hemorrhaged to a mean arterial pressure of 30 mmHg for 90 min, but they were not resuscitated. Alveolar liquid clearance was measured by the concentration of labeled and unlabeled protein over 2 h in an isosmolar physiological solution of 5% albumin that had been instilled into one lung. Hemorrhaged rats developed a severe metabolic acidosis that was associated with a 5- to 10-fold rise in plasma epinephrine levels. There was a 60% increase in alveolar liquid clearance in the hemorrhaged rats compared with control rats (55 +/- 6 vs. 34 +/- 7%; P < 0.05). Amiloride (10(-4) M) or propranolol (10(-4)M) inhibited the increase in alveolar liquid clearance. Thus the endogenous release of catecholamines associated with hemorrhagic shock markedly stimulates alveolar fluid clearance by a beta-adrenergic-mediated stimulation of active sodium transport. These data suggest a new, previously unrecognized mechanism that may protect against alveolar flooding in the acute phase of hemorrhagic shock.     

Muscarinic receptors in rat cortex, hippocampus, hypothalamus and brainstem following transient forebrain ischemia and hemorrhagic shock

[3H]Quinuclidinyl benzilate binding properties of cerebral cortex, hippocampus, hypothalamus and brainstem of rats subjected to transient forebrain ischemia or severe hemorrhagic shock were investigated. Maximal binding capacities (Bmax) were not significantly different from control animals in either model. On the other hand, significant increases in binding affinities at all four brain regions in the ischemia-reperfusion group and at hypothalamic and brainstem membranes in the hemorrhagic shock group were observed. Kd values obtained in cortex and hippocampus of animals in shock were similar to control values. It was concluded that in brain ischemia models, the number of brain muscarinic receptors do not change at early stages, but binding affinities increase most likely due to systemic hypotension rather than reperfusion. The well-developed circle of Willis seems to protect cortical and hippocampal muscarinic receptors from hypoxia-induced changes.     

Regulation and functional significance of phospholipase D in myocardium

There is now clear evidence that receptor-dependent phospholipase D is present in myocardium. This novel signal transduction pathway provides an alternative source of 1,2-diacylglycerol, which activates isoforms of protein kinase C. The members of the protein kinase C family respond differently to various combinations of Ca2+, phosphatidylserine, molecular species of 1,2-diacylglycerol and other membrane phospholipid metabolites including free fatty acids. Protein kinase C isozymes are responsible for phosphorylation of specific cardiac substrate proteins that may be involved in regulation of cardiac contractility, hypertrophic growth, gene expression, ischemic preconditioning and electrophysiological changes. The initial product of phospholipase D, phosphatidic acid, may also have a second messenger role. As in other tissues, the question how the activity of phospholipase D is controlled by agonists in myocardium is controversial. Agonists, such as endothelin-1, atrial natriuretic factor and angiotensin II that are shown to activate phospholipase D, also potently stimulate phospholipase C-beta in myocardium. PMA stimulation of protein kinase C inactivates phospholipase C and strongly activates phospholipase D and this is probably a major mechanism by which agonists that promote phosphatidyl-4,5-bisphosphate hydrolysis secondary activate phosphatidylcholine-hydrolysis. On the other hand, one group has postulated that formation of phosphatidic acid secondary activates phosphatidyl-4,5-bisphosphate hydrolysis in cardiomyocytes. Whether GTP-binding proteins directly control phospholipase D is not clearly established in myocardium. Phospholipase D activation may also be mediated by an increase in cytosolic free Ca2+ or by tyrosine-phosphorylation.     

Plasma endotoxin and cytokine concentrations in patients with hemorrhagic shock

OBJECTIVES: The roles of cytokines and endotoxin in hemorrhagic shock, particularly the translocation of endotoxin and bacteria during hemorrhagic shock, were investigated. DESIGN: Prospective study. SETTING: Critical care and emergency center of a university hospital. PATIENTS: Twenty-nine patients with hemorrhagic shock and 20 healthy controls. INTERVENTIONS: Serial blood samples were collected from both study and control patients. Standard resuscitation techniques were used. MEASUREMENTS AND MAIN RESULTS: Plasma levels of endotoxin and various cytokines were determined repeatedly during hemorrhagic shock. Endotoxin was measured using an endotoxin-specific assay in addition to a new perchloric acid method for pretreatment of plasma. Cytokines were measured by commercial enzyme-linked immunosorbent assays. Plasma endotoxin concentrations remained within the normal range for 7 days after admission. Although levels of tumor necrosis factor-alpha and several interleukins increased slightly in some patients, these cytokines did not reach the levels seen in septic shock. CONCLUSIONS: Translocation of bacteria or endotoxin from the gastrointestinal tract into the bloodstream has been noted in animal experiments; however, translocation was not detected in our patients with hemorrhagic shock.     

Quantitative cytochemical changes induced by dexamethasone and adrenalectomy in rat liver chromatin

A wild-type strain of Staphylococcus aureus, which inactivates a wide variety of aminoglycosides (except the gentamicin components), has been found to harbor a plasmid (RAp01) that mediates the biosynthesis of a nucleotidyltransferase. This enzyme modifies the 4'-hydroxy function of these antibiotics. The plasmid has been studied, the enzyme responsible for this resistance pattern has been isolated by affinity chromatography, and its kinetics and physicochemistry have been characterized. The target of this enzyme has also been located by demonstrating the structure of one inactivated compound, 4'-(O)-adenylyltobramycin.     

Involvement of nitric oxide in hyporeactivity of rat mesenteric vascular bed during endotoxic shock: effect of dexamethasone and endothelin-I

The present study was carried out on mesenteric vascular bed from LPS-injected rats in order to investigate the cause of hyporesponsiveness in resistance blood vessels, during septic shock syndrome. The involvement of L-Arg/NO pathway was evaluated by administration of L-Arg, which produced a decrease in perfusion pressure in LPS-treated rats, whereas it was ineffective in control rats. Furthermore, DEX-pretreatment in endotoxaemic rats significantly reduced the vasorelaxation by L-Arg, whereas it was ineffective to reverse vascular hyporeactivity occurring in septic shock. In order to evaluate whether hyporesponsiveness could be due to defects in contraction mechanisms, we tested the effect of ET-I. This peptide was able to markedly enhance the contractile response to NA in LPS-treated rats. Our findings suggest that vascular hyporesponsiveness during septic shock may depend on both activation of the L-Arg/NO pathway and alterations in post-receptor mechanisms involving calcium handling.     

Serotonin is involved in the ACTH-induced reversal of hemorrhagic shock in anesthetized rats

In a rat model of volume-controlled hemorrhagic shock (mean arterial pressure = 20-24 mm Hg) causing the death of all saline-treated animals within 30 min, the i.v. bolus injection of ACTH-(1-24) (160 micrograms/kg) produced an almost complete and sustained reversal of the shock condition, with recovery of arterial blood pressure, pulse pressure and respiratory rate, and with 100% survival at the end of the experiment (2 h). The serotonin-depleting agent p-chlorophenylalanine (316 mg/kg i.p., administered 66-70 h before hemorrhage) almost completely prevented the effect of ACTH. The 5-HT1/5-HT2 receptor antagonist, methysergide, prevented the effect of ACTH completely when injected i.v. (5 mg/kg), but only in part when injected into a brain ventricle (i.c.v.) (15 micrograms/rat); the 5-HT2 antagonist, ketanserin, prevented the effect of ACTH completely when injected i.c.v. (1.5 micrograms/rat), but only in part when injected i.v. (0.5 mg/kg); the 5-HT3 antagonist, MDL 72222, largely prevented the effect of ACTH when injected i.c.v. (10 micrograms/rat), but had no influence at all when injected i.v. (3 mg/kg); finally, the 5-HT4 antagonist, GR 125487, had no effect when injected i.v. (5 micrograms/kg) or when injected i.c.v. (30 ng/rat). Overall, these data indicate that both CNS and peripheral serotonin play an important role in the complex mechanism of the ACTH-induced hemorrhagic shock reversal.     

Anisotropy of water diffusion in the myocardium of the rat

Pulsed field gradient nuclear magnetic resonance methods combined with nuclear magnetic resonance imaging were used to determine the water diffusion anisotropy in perfused rat hearts at 37 degrees C. It was found that the observed diffusion coefficient D(app) (apparent diffusion coefficient) depends on the orientation of the applied gradient g. When g is parallel to the epicardial surface, the observed diffusivity is D(app) parallel = 1.8 +/- 0.4 x 10(-9) m2.s-1, whereas when g is perpendicular to it, diffusivity is D(app) perpendicular = 2.5 +/- 0.5 x 10(-9) m2.s-1. To better characterize this directional dependence, images of the second-order diffusion tensor D of the myocardium were obtained. These data demonstrate several essential features of cardiac myoarchitecture, including the helicity of fiber orientation with respect to the ventricular axis and the variation of fiber pitch angle with transmural depth. Diffusion anisotropy may be quantified in a coordinate-independent manner by the eigenvalues of the diffusion tensor. In the myocardial midwall, these eigenvalues were E1 = 3.29 +/- 0.57, E2 = 2.01 +/- 0.42, and E3 = 0.77 +/- 0.58 x 10(-9) m2.s-1 (mean +/- SD). These data suggest that myocardial water diffusion is essentially unrestricted parallel to the myofibers. They further show that failure to measure the complete diffusion tensor may lead to substantial underestimates of diffusion anisotropy in the myocardium.