Biomedical subjects

P D Harris

Publications and source records attributed to P D Harris.

At least 37 records · Page 2Linked to original sources

PubMed2000

Sustained infection induces 2 distinct microvascular mechanisms in the splanchnic circulation.

BACKGROUND: Altered intestinal blood flow during systemic inflammation leads to organ dysfunction. Mucosal ischemia occurs during sepsis despite an increase in portal blood flow. We hypothesized that separate mechanisms are active in the large resistance and small mucosal microvessels to account for this dichotomy. METHODS: Chronic infection was induced in rats by bacterial inoculation (Escherichia coli and Bacteroides fragilis) of an implanted subcutaneous sponge. Separate groups were studied at 24 and 72 hours after a single inoculation of bacterium or 24 hours after a second inoculation (ie, 72 hours of sepsis). Time-matched controls were used for each group. Intravital microscopy of the terminal ileum was used to assess endothelial-dependent vasodilation to acetylcholine (10(-9) to 10(-5) mol/L) in resistance (A(1)) and premucosal (A(3)) arterioles. Threshold sensitivity (-log of 20% response dose) was calculated from dose response curves for each animal. RESULTS: Vasodilator sensitivity to acetylcholine in A(1) arterioles was significantly decreased at 24 hours, and these changes persisted up to 72 hours after a single bacterial inoculation. There was no change in the dilator sensitivity of A(3) arterioles after a single inoculation. When there was a challenge with a second bacterial inoculation, there was a reversal of the A(1) dilator response and an increase in A(3) sensitivity. CONCLUSIONS: An initial septic event results in a decrease in dilator reactivity in the resistance A1 arterioles that persists for at least 72 hours. A sustained septic challenge results in increased dilator reactivity in both A(1) and A(3) vessels. This enhanced sensitivity during sepsis suggests that more than 1 therapeutic approach to preservation of intestinal blood flow will be necessary.

Acetylcholine↗

PubMed2000

Intestinal helminths of spiny mice (Acomys cahirinus dimidiatus) from St Katherine's Protectorate in the Sinai, Egypt.

Spiny mice, Acomys cahirinus dimidiatus, inhabiting the wadis close to St Katherine in the mountains of the Sinai peninsula, were trapped and their helminth parasites were studied. Sixty one mice provided faeces for analysis and 27 were killed and autopsied. Six species of helminths were recorded (the spirurid nematodes, Protospirura muricola (74.1%) and Mastophorus muris (11.1%), the oxyuroid nematodes, Dentostomella kuntzi (59.3%), Aspiculuris africana (3.7%), and Syphacia minuta (3.7%) and the hymenolepidid cestode Rodentolepis negevi (18.5%)). The spirurids were the dominant species present, accounting for up to 0.87% of total host body weight. Analysis of worm weights and lengths suggested that transmission had been taking place in the months preceding our study. No sex difference in the prevalence or abundance of spirurids was detected. Significant differences were identified in the abundance of total nematode burdens and the mean helminth species richness between the three wadis which provided multiple captures of mice. There was also a marked effect of host age on both parameters. A highly significant positive correlation between spirurid egg counts and total worm biomass indicated that non-invasive techniques based on egg counts could be used to quantify worm burdens and when this technique was applied to a larger sample size (n = 61), a significant difference between sites but no host sex or age effects were detected for spirurid faecal egg counts. The data suggest that there are differences between helminth component communities infecting spiny mice in different neighbouring wadis, a hypothesis which will be explored further through our continuing studies in the Sinai.

Age Factors↗

PubMed1999

Decreased alpha-adrenergic response in the intestinal microcirculation after "two-hit" hemorrhage/resuscitation and bacteremia.

BACKGROUND: The two-hit theory of multiple organ dysfunction syndrome proposes that an initial insult primes the host for an altered response to subsequent stimuli. We have previously documented enhanced dilator tone in the small intestine after a two-hit insult; however, the effects on vasoconstrictor function are unknown. We postulated that prior hemorrhage and resuscitation followed by bacteremia would alter microvascular responsiveness to alpha-adrenergic stimulation. METHODS: Male Sprague-Dawley rats underwent fixed-volume hemorrhage with resuscitation (H/R) or sham procedure (Sham). At 24 or 72 h, in vivo videomicroscopy of the small intestine was performed (inflow A1 and premucosal A3 arterioles). Constrictor function was assessed by topical application of norepinephrine (NE; 10(-8)-10(-6) M) before and 1 h after intravenous Escherichia coli or saline. RESULTS: Sham, 24 or 72 h H/R, and E. coli alone produced no significant changes in A1 or A3 response to NE. Sequential H/R + E. coli resulted in decreased constrictor response in both A1 (72 h H/R + E. coli-38% from baseline vs Sham - 54%, P < 0.05) and A3 arterioles (-8% vs -51%, P < 0.05) at high doses of NE (10(-6) M). CONCLUSIONS: Prior H/R primes the intestinal microvasculature for an altered response during a subsequent stress and these effects persist for up to 72 h following H/R. Sequential insults in this two-hit model caused marked hyporesponsiveness to NE. These alterations in control of microvascular tone might contribute to the hemodynamic compromise of sepsis, impair mucosal blood flow, and contribute to the development of MODS.

Adrenergic alpha-Agonists↗

PubMed1999

Glucose and glutamine gavage increase portal vein nitric oxide metabolite levels via adenosine A2b activation.

INTRODUCTION: Postprandial intestinal hyperemia is a complex vascular response during nutrient absorption. Many mediators have been studied including enteric reflexes, GI hormones, and absorption-stimulated metabolic mediators such as pH and adenosine. We have shown that nitric oxide (NO) mediates premucosal arteriolar dilation during glucose absorption and that glucose-induced portal vein NO metabolite production requires adenosine A2b receptor activation. We hypothesize that Na+-linked absorption of l-glutamine or l-glycine might also stimulate NO release in the enteroportal circulation via adenosine A2b receptors. METHODS: Male Sprague-Dawley rats (190-220 g) were anesthetized with urethane/alpha-chloralose and cannulated for hemodynamic monitoring and blood sampling. A right paramedian abdominal incision was made for access to both the stomach (gavage) and the portal vein (blood sampling). Animals received intragastric nutrient gavage (saline, d-glucose, l-glutamine, racemic glycine, or oleic acid) with and without adenosine A2b receptor blockade. NO metabolites (NOx) were measured by a fluorescent modified-Greiss assay at baseline and 30 min after nutrient gavage. RESULTS: Glucose and glutamine gavage increased portal NOx levels compared to baseline, while glycine and oleic acid gavage did not. Adenosine A2b antagonism returned NOx levels to baseline in both glucose and glutamine gavage animals, but did not alter portal NOx levels in glycine- or oleic acid-treated animals. CONCLUSIONS: These data suggest that nutrient-induced adenosine is involved in a signaling process from the intestinal epithelium to nitric oxide-producing cells elsewhere in the vasculature. Adenosine A2b receptors are required for NO production during Na+-linked glucose or glutamine absorption.

Administration, Oral↗

PubMed1999

Subacute sepsis impairs vascular smooth muscle contractile machinery and alters vasoconstrictor and dilator mechanisms.

INTRODUCTION: Sepsis results in hyporesponsiveness to alpha-adrenergic stimulation. This is thought to be mediated by the release of vasoactive compounds from the septic endothelium or by the direct effect of sepsis on vascular smooth muscle (VSM) contractile mechanics and machinery. Previous studies have used lethal models of sepsis or endotoxemia to examine this phenomenon. The present study utilizes a clinically relevant, nonlethal model of soft tissue infection to determine the effects of sepsis on alpha-adrenergic mechanisms. We hypothesize that subacute sepsis causes impaired alpha-adrenergic vascular responsiveness by a combination of effects on adrenergic constrictor mechanisms, endogenous dilator tone, and VSM contractile function. METHODS: Male Sprague-Dawley rats underwent implantation of a 2 x 2-cm2 gauze sponge into a subcutaneous pocket created at the base of the tail. Five days after implantation, sepsis (S) was induced by inoculation of the sponge with 10(9) CFU Escherichia coli and Bacteroides fragilis. Controls (C) were inoculated with saline. Thoracic aortic harvest was performed 24 and 48 h after sponge inoculation for organ bath ring studies. Receptor-mediated (phenylephrine) and nonreceptor-mediated (KCl) maximum force of contraction (Fmax) was measured. Vessel sensitivity (pD2) to phenylephrine, acetylcholine, and KCl was calculated from dose-response curves. RESULTS: At 24 h, sepsis resulted in a lower Fmax to phenylephrine (1.15 for C vs 0.5 for S, P < 0.05 by ANOVA), despite an increase in vessel sensitivity (pD2) to alpha-adrenergic stimulation (6.70 for C vs 6.88 for S, P < 0.05 by ANOVA). Fmax to KCl was lower in septic animals at 24 h (3. 50 for C vs 2.77 for S, P < 0.05 by ANOVA) and sensitivity to acetylcholine (pD2) was markedly increased (6.56 for C vs 7.23 for S, P < 0.05 by ANOVA). At 48 h, the impairment in Fmax to alpha-adrenergic stimulation (2.29 for C vs 1.72 for S, P < 0.05 by ANOVA) and KCl (3.5 for C vs 3.08 for S. P < 0.05 vs 24 h C by ANOVA) persisted without any change in sensitivity to phenylephrine or acetylcholine. CONCLUSIONS: Subacute sepsis results in an early suppression of maximum contractile force despite an increase in adrenergic receptor sensitivity (pD2). This may be secondary to an elevation in dilator sensitivity combined with a direct effect of sepsis on VSM contractile mechanisms. Later in the septic process, however, alpha-adrenergic hyporesponsiveness ( downward arrow Fmax) is primarily due to changes in VSM contractile machinery.

Acetylcholine↗

PubMed1999

Altered vasoconstrictor and dilator responses after a "two-hit" model of sequential hemorrhage and bacteremia.

BACKGROUND: The "two-hit" theory of multiple organ dysfunction (MOD) proposes that an initial insult, such as hemorrhage (HEM), primes the host for an abnormal response to a second stress such as infection. The immunologic/inflammatory component of this theory has been well examined; however, the effects on vascular responsiveness are poorly understood. We hypothesized that HEM primes the vasculature for an altered response to a second pathophysiologic stress. METHODS: Male Sprague-Dawley rats underwent a fixed-volume HEM with resuscitation (H/R) or sham procedure (Sham). At 48 h, animals were given iv E. coli or saline and followed for 1 h. Thoracic aortic rings were then placed in organ baths containing Krebs buffer aerated with 95% O2, 5% CO2. Cumulative dose-response curves to phenylephrine (PHE) and acetylcholine (ACH) were obtained. Maximum force of contraction (Fmax) was measured and pD2 values (receptor sensitivity) were derived. RESULTS: H/R alone resulted in heightened constrictor tone and blunted dilator tone. E. coli reduced Fmax in response to PHE by 50% in Sham vs 76% in H/R. Receptor sensitivity (pD2) to PHE was reduced to a greater degree in H/R (3-fold vs 2-fold). These animals also had a more pronounced enhancement of ACH receptor sensitivity (7-fold vs 2-fold). CONCLUSIONS: Hemorrhage primes the vasculature for an altered response to a subsequent stress. When infection is added as a "second hit," responsiveness to adrenergic agents is diminished and dilator tone is increased. These data may explain the cardiovascular derangements seen clinically in patients who develop MODS after major hemorrhage followed sequentially by infection.

Acetylcholine↗

PubMed1999

The susceptibility of Atlantic salmon (Salmo salar L.) x brown trout (Salmo trutta L.) hybrids to Gyrodactylus salaris Malmberg and Gyrodactylus derjavini Mikailov.

Salmo salar and Salmo trutta co-exist in coastal river systems in Europe and produce hybrids with little loss of viability or growth. This report describes the susceptibility of pure full-sibs of S. salar and S. trutta and their reciprocal half-sib hybrids to their respective gyrodactylids, Gyrodactylus salaris and Gyrodactylus derjavini. The pure-bred salmon and trout, and half-sib hybrids, were produced using eggs and sperm from wild anadromous S. salar (River Alta stock, North Norway) and wild anadromous S. trutta (River Fossbekk stock, Southwest Norway). Infections were initiated by exposing experimental fishes (0+) to S. salar naturally infected with G. salaris (River Lierelva strain) or S. trutta naturally infected with G. derjavini (River Sandvikselva strain). Fishes were then kept individually isolated under standardized conditions at 12 degrees C. Pure-bred S. salar were susceptible but frequently mounted a response to G. salaris without eliminating the infection, whereas pure-bred S. trutta were innately resistant to this species. Pure-bred S. trutta ranged from innately resistant to susceptible to G. derjavini but later most of the susceptible trout mounted a host response to G. derjavini. Pure-bred S. salar were also susceptible to this species, although parasite population growth rates were reduced and a host response frequently appeared eliminating G. derjavini. The abundance of both gyrodactylids was lower on the hybrids than on their respective pure-bred natural hosts, and a parental sire- and dam-influence on the resistance of hybrids was observed. When the sire was S. salar, the susceptibility of hybrids to G. salaris was similar to that of pure S. trutta; when the dam was S. salar both innately resistant, intermediately susceptible and responding individuals were present. In the case of G. derjavini, when the sire was S. trutta, infections on hybrids were similar to those on pure S. salar; when the dam was S. trutta, an increased level of susceptibility was observed. The present results provide evidence that: (1) Norwegian salmon stocks are variable in their susceptibility/resistance, with some fish able to control S. salaris infections; (2) trout stocks are innately resistant to G. salaris; (3) individual trout show a spectrum in susceptibility/resistance to G. derjavini, ranging from innate resistance through slightly susceptible to highly susceptible but with acquired resistance controlling infection; (4) although G. derjavini infections grow poorly on salmon, this host stock is susceptible to the parasite, but can limit infection by a host reaction; (5) susceptibility/resistance traits to gyrodactylids are genetically controlled and resistance can be transferred as a dominant trait through interspecific crosses between different salmonids; (6) interspecific hybrids between susceptible and resistant salmonids have a pattern of susceptibility to gyrodactylids intermediate to that of the parents; (7) resistance to gyrodactylids may be controlled by relatively few genes in salmonids; (8) epidemiologically, hybrids may act as a reservoir for gyrodactylids, may support a wider diversity of species than either parent and may disseminate gyrodactylids of both host species.

Animals↗

PubMed1999

Combined ribosomal DNA and morphological analysis of individual gyrodactylid monogeneans.

A method is presented for the isolation and analysis of hamuli, marginal hooks, and bars from individual gyrodactylid monogeneans using scanning electron microscopy (SEM), while simultaneously processing parasites for rDNA analysis using the polymerase chain reaction (PCR). The haptors of ethanol-fixed gyrodactylids were protease digested to liberate hooks for SEM, whereas DNA extracted from the bodies was used for PCR. The method resulted in hooks and hamuli being prepared from more than 90% of Gyrodactylus turnbulli individuals, a significant improvement on previously published digestion-based SEM techniques. PCR on the same parasites was less successful, but sequence data were obtained from 50% of individuals. Amplification of rDNA internal-transcribed spacer regions from individual worms used for SEM gave PCR products consistent with those predicted from our previous sequence analysis. This method allows the correlation of morphology and DNA sequence from the same individual and can be applied to ethanol-fixed material, such as field collected and museum specimens.

Animals↗

PubMed1998

Life history specializations of monogenean flatworms: a review of experimental and microscopical studies.

This review illustrates the use of experimental approaches combined with microscopy to study the biology of monogenean parasites. Studies of feeding, development, reproduction, and systematics have been based on gyrodactylids, flatworms infecting teleost fishes. In a contrasting system involving an amphibian host in a desert environment, analysis of adaptations to extreme conditions has focused on Pseudodiplorchis americanus. The unusual reproductive strategies, particularly the interactions between mother and offspring, are highlighted for both monogeneans. Species of Gyrodactylus are viviparous, maintaining up to three generations of embryos simultaneously in utero, and many of their reproductive specializations are related to progenesis. Embryo nutrition takes place via a metabolically-active syncytial uterine lining that has close association with the parental gut. Microscopy has also proved an essential adjunct to molecular studies of speciation and host specificity. P. americanus is ovoviviparous and the adaptations for embryo maintenance are unique. The primary keratin-type eggshell is replaced by a flexible secondary elastin capsule produced by the uterus; parental nutrients are transferred through cytoplasmic connections to the developing embryo. TEM has demonstrated unique adaptations of P. americanus to its micro-environments, including secretion of tegumental vesicles that provide protection from digestive enzymes during migration through the host gut. This paper highlights the potential of monogeneans for studies of fundamental biological principles.

Animals↗

PubMed1998

Small intestinal production of nitric oxide is decreased following resuscitated hemorrhage.

BACKGROUND: Small intestine microvascular vasoconstriction and hypoperfusion develop after resuscitation (RES) from hemorrhage (HEM), despite restoration of central hemodynamics. The responsible mechanisms are unclear. We hypothesized that the microvascular impairment following HEM/RES was due to decreased intestinal microvascular nitric oxide (NO) production. METHODS: Male Sprague-Dawley rats (195-230 g) were utilized and three experimental groups were studied: (1) SHAM (cannulated but no HEM), (2) HEM only, and (3) HEM/RES. HEM was to 50% of baseline mean arterial pressure for 60 min, and RES was with shed blood and an equivalent volume of saline. Ex vivo isolated intestinal perfusion and a fluorometric modification of the Greiss reaction were used to quantify production of NO metabolites (NOx). Perfusate von Willebrand factor (vWF) was used as an indirect marker of endothelial cell activation or injury. To assess the degree of NO scavenging by oxygen-derived free radicals, immunohistochemistry was used to detect nitrotyrosine formation in the intestine. RESULTS: Intestinal NOx decreased following HEM/RES (SHAM 1.35 +/- 0.2 mM vs HEM/RES 0.60 +/- 0.1 mM, P < 0.05), but not with HEM alone (1.09 +/- 0.3 mM). There were no differences in serum NOx levels between the three groups. Release of vWF was increased during the HEM period (SHAM 0.18 +/- 0.1 g/dl vs HEM 1.66 +/- 0.6 g/dl, P < 0.05). There was no detectable nitrotyrosine formation in any group. CONCLUSIONS: Intestinal NO metabolites decrease following HEM/RES. Elevated vWF levels during HEM and the lack of detectable nitrotyrosine suggest that this is due to decreased endothelial cell production of NO. HEM/RES-induced endothelial cell dysfunction may contribute to persistent small intestine post-RES hypoperfusion and vasoconstriction.

Animals↗

PubMed1998

Ecological and genetic evidence for clonal reproduction in Gyrodactylus gasterostei Glaser, 1974.

Reproduction of isolated Gyrodactylus gasterostei from the three-spined stickleback, Gasterosteus aculeatus, has been investigated. Three lineages of isolated parasites, without opportunity for cross- or self-fertilisation, continued to reproduce for up to 19 generations (224 births) until experiments were discontinued. Most parasites gave birth to two daughters only, although four individuals in one lineage gave birth three times. After the final birth the empty uterus expanded, while a large oocyte remained in the seminal receptacle and appeared capable of development. Reproduction by isolated flukes was further analysed using quantitative genetic techniques, applied to the hamuli and marginal hooks. The variance of hamulus and marginal hook dimensions in lineages of isolated flukes (which, if sexual must be highly inbred) was similar to that observed in natural populations. Crude heritability appeared high, but a significant relationship between hook size and environmental temperature was noted, and when this was controlled for, the heritability of all characters became negligible. Genetic drift was not observed over 17 generations and variance within isolated lines accounted for almost all of the total variance. Genes controlling hamulus or marginal hook dimensions did not progress to fixation, although the number of generations (more than 10) was sufficient to ensure homozygosity if self-fertilisation were occurring. These observations indicate that reproduction by isolated G. gasterostei continues indefinitely, but does not involve sex. The limited variance of natural populations suggests that these are also clonal with observed variation being largely environmental. Sexual reproduction may therefore be rare in G. gasterostei.

Animals↗

PubMed1998

Killing of Gyrodactylus salaris (Platyhelminthes, Monogenea) mediated by host complement.

Gyrodactylus salaris, an important pathogen of Atlantic salmon Salmo salar, has been shown to be highly sensitive to factors in host serum and mucus, being killed rapidly (50% within 1 h) by serum at a dilution of 1:200. The time needed for killing was inversely proportional to serum concentration. Similar effects were noted using host mucus, which contained approximately 1/20th of the anti-Gyrodactylus activity of serum. Serum activity was abolished completely by heating at 45 degrees C for 30 min, and by addition of EDTA, but not by EGTA + 1 mM magnesium ions. Activity was not dependent on whether the serum was from infected or naive fishes, nor was it species specific. Attempts to pre-coat parasites in salmon anti-Gyrodactylus antibodies also failed to enhance the activity of fresh serum. These observations suggest that killing is due to the complement system of the host, acting via the alternate pathway. G. salaris appears to be exceptionally sensitive to complement, being killed at concentrations which could be experienced in vivo. The role of complement in the protection of fishes against gyrodactylid infection therefore deserves further investigation.

Animals↗

PubMed1998

Complement inhibition prevents gut ischemia and endothelial cell dysfunction after hemorrhage/resuscitation.

BACKGROUND: Complement, a nonspecific immune response, is activated during hemorrhage/resuscitation (HEM/RES) and is involved in cellular damage. We hypothesized that activated complement injures endothelial cells (ETCs) and is responsible for intestinal microvascular hypoperfusion after HEM/RES. METHODS: Four groups of rats were studied by in vivo videomicroscopy of the intestine: SHAM, HEM/RES, HEM/RES + sCR1 (complement inhibitor, 15 mg/kg intravenously given before resuscitation), and SHAM + sCR1. Hemorrhage was to 50% of mean arterial pressure for 60 minutes followed by resuscitation with shed blood plus an equal volume of saline. ETC function was assessed by response to acetylcholine. RESULTS: Resuscitation restored central hemodynamics to baseline after hemorrhage. After resuscitation, inflow A1 and premucosal A3 arterioles progressively constricted (-24% and -29% change from baseline, respectively), mucosal blood flow was reduced, and ETC function was impaired. Complement inhibition prevented postresuscitation vasoconstriction and gut ischemia. This protective effect appeared to involve preservation of ETC function in the A3 vessels (SHAM 76% of maximal dilation, HEM/RES 61%, HEM/RES + sCR1 74%, P < .05). CONCLUSIONS: Complement inhibition preserved ETC function after HEM/RES and maintained gut perfusion. Inhibition of complement activation before resuscitation may be a useful adjunct in patients experiencing major hemorrhage and might prevent the sequelae of gut ischemia.

Acetylcholine↗

PubMed1998

Microvascular changes explain the "two-hit" theory of multiple organ failure.

OBJECTIVE: The objective was to determine intestinal microvascular endothelial cell control after sequential hemorrhage and bacteremia. SUMMARY BACKGROUND DATA: Sepsis that follows severe hemorrhagic shock often results in multiple system organ failure (MSOF) and death. The sequential nature of this clinical scenario has led to the idea of a "two-hit" theory for the development of MSOF, the hallmark of which is peripheral vasodilation and acidosis. Acute bacteremia alone results in persistent intestinal vasoconstriction and mucosal hypoperfusion. Little experimental data exist to support the pathogenesis of vascular dysregulation during sequential physiologic insults. We postulate that hemorrhagic shock followed by bacteremia results in altered microvascular endothelial cell control of dilation and blood flow. METHODS: Rats underwent volume hemorrhage and resuscitation. A sham group underwent the vascular cannulation without hemorrhage and resuscitation, and controls had no surgical manipulation. After 24 and 72 hours, the small intestine microcirculation was visualized by in vivo videomicroscopy. Mean arterial pressure, heart rate, arteriolar diameters, and A1 flow by Doppler velocimetry were measured. Endothelial-dependent dilator function was determined by the topical application of acetylcholine (ACh). After 1 hour of Escherichia coil bacteremia, ACh dose responses were again measured. Topical nitroprusside was then applied to assess direct smooth muscle dilation (endothelial-independent dilator function) in all groups. Vascular reactivity to ACh was compared among the groups. RESULTS: Acute bacteremia, with or without prior hemorrhage, caused significant large-caliber A1 arteriolar constriction with a concomitant decrease in blood flow. This constriction was blunted at 24 hours after hemorrhage but was restored to control values by 72 hours. There was a reversal of the response to bacteremia in the premucosal A3 vessels, with a marked dilation both at 24 and 72 hours. The sequence of hemorrhage and E. coli resulted in a progressive enhanced reactivity to the endothelial-dependent stimulus of ACh in the A3 vessels at 24 and 72 hours. Reactivity to endothelial-independent smooth muscle relaxation and subsequent vessel dilation was similar for all groups. CONCLUSIONS: These data indicate that there is altered endothelial control of the intestinal microvasculature after hemorrhage in favor of enhanced dilator mechanisms in premucosal vessels with enhanced constrictor forces in inflow vessels. This enhanced dilator sensitivity is most evident in small premucosal vessels. This experimental finding supports the premise that an initial pathophysiologic stress alters the subsequent microvascular blood flow responses to systemic inflammation. These changes in the intestinal microcirculation are in concert with the "two-hit" theory for MSOF.

Acute Disease↗

PubMed1998

Selective microvascular endothelial cell dysfunction in the small intestine following resuscitated hemorrhagic shock.

Following resuscitation (RES) from hemorrhagic shock (HEM), intestinal microvessels develop progressive vasoconstriction that impairs mucosal blood flow, despite central hemodynamic RES. These events might have clinical consequences secondary to occult intestinal ischemia. We hypothesized that the microvascular impairments were due to progressive endothelial cell dysfunction and an associated reduction in the dilator, nitric oxide (NO), following HEM/RES. Male Sprague-Dawley rats, were monitored for central hemodynamics and the terminal ileum was studied with in vivo videomicroscopy. HEM was 50% of baseline mean arterial pressure (MAP) for 60 min, and RES was with shed blood + 1 volume of normal saline (NS). Following HEM/RES, acetylcholine (10)(-7), 10(-5) M) was topically applied and ileal inflow (A1) and premucosal arteriolar diameters were measured to assess endothelial-cell function at 60 and 120 min post-RES. Normalization of MAP, cardiac output, and heart rate demonstrated adequate systemic resuscitation. Post-RES vasoconstriction developed in A1 (-25%) and premucosal (-28%) arterioles with an associated reduction in A1 flow (-47%). However, there was a selective impairment of endothelial-dependent dilation that was manifested only in the smaller premucosal arterioles and not in the inflow, A1 arterioles. This suggests that multiple mechanisms are involved in the development of the post-RES vasoconstriction. The premucosal response was likely mediated by endothelial cell dysfunction, while the A1 response was probably the result of enhanced vasoconstrictor forces. This early microvascular dysfunction might contribute to the late sequelae of intestinal ischemia and might alter microvascular responses to subsequent systemic insults.

Acetylcholine↗

PubMed1998

E. Coli bacteremia-induced changes in the skeletal muscle microcirculation vary with anesthetics.

AIM: To test if anesthetic procedures change the hemodynamic pattern in animals with experimental septic shock. METHODS: The effect of two anesthetics on systemic hemodynamic and skeletal muscle microcirculatory responses in high cardiac output live E. coli bacteremia was studied in rats and compared to the effect of two other anesthetic procedures in previously published studies. RESULTS: Baseline blood pressures and cardiac outputs were similar in rats with decerebrate, ketamine/xylazine, pentobarbital or urethane/chloralose anesthesia. There was a relative baseline tachycardia in decerebrate rats. Ketamine/xylazine anesthetized rats had reduced blood pressure, cardiac output, and heart rate. In decerebrate, pentobarbital, and urethane/chloralose anesthesia, cardiac output increased initially during bacteremia but did not remain elevated in pentobarbital anesthesia. Blood pressure and heart rate remained constant in pentobarbital, decerebrate, and urethane/chloralose anesth esia. During bacteremia, cardiac output, blood pressure, and vascular resistance did not change with ketamine/xylazine, but the heart rate increased. Baseline diameters of cremaster muscle large (A1) arterioles were higher in decerebrate anesthesia. A1 arterioles constricted during high cardiac output bacteremia in decerebrate rats, and pentobarbital or urethane/chloralose-anesthetized rats. A4 arterioles in bacteremia dilated in decerebrate and pentobarbital anesthesia, but did not change under urethane/chloralose and ketamine/xylazine anesthesia. CONCLUSION: Anesthetics influence baseline systemic variables and the response of systemic hemodynamics of rats to E. coli bacteremia. During bacteremia, anesthetics primarily affect the reactivity of skeletal muscle small arterioles. Ketamine/xylazine anesthesia has the most pronounced effect on systemic and microcirculatory variables and seems to be an inappropriate choice in sepsis experiments in rats.

Anesthetics↗

PubMed1997

Glucose-induced intestinal hyperemia is mediated by nitric oxide.

UNLABELLED: Glucose-induced absorptive hyperemia of the intestine has been well demonstrated through microsphere blood flow experiments. We have previously demonstrated that glucose, when applied topically to rat ileal epithelium, restores microvascular vessel diameters and blood flow following Escherichia coli bacteremia or hemorrhage/resuscitation. However, the mechanisms of this hyperemia are not completely understood. We hypothesize that nitric oxide is a mediator of the microvascular response to glucose exposure on the rat intestinal epithelium. METHODS: Male Sprague-Dawley rats, 200-225 g, were monitored for hemodynamic stability with mean arterial blood pressure and heart rate. A 2-cm segment of the terminal ileum with intact neurovascular supply was exposed for intravital videomicroscopy. Intestinal arteriolar diameters (A1D, inflow; and A3D, premucosal arterioles) and microvascular blood flow (A1Q) were measured following topical application of isoosmotic glucose or saline, with or without l-NAME (LN, 100 mM), a competitive inhibitor of nitric oxide synthase. Statistical analysis was performed by ANOVA followed by Tukey-Kramer honestly significant difference test. RESULTS: All data are expressed as mean percentage changes from baseline +/- standard error of the mean. Hemodynamic variables did not change during the experimental procedure and there were no significant differences among group baselines. Addition of isotonic glucose to the bath solution caused a significant increase in A3D that persisted throughout the experiment (at 30 min, 19.2 +/- 4.2 vs -3.9 +/- 4.5, P < 0.05). This vasodilation was blocked by topical administration of LN (3.1 +/- 2.9, P < 0.05). A1D remained at baseline levels (saline and glucose) or constricted (LN) in all groups. Topical LN also attenuated A1Q in both the saline and glucose groups. CONCLUSIONS: These data demonstrate that glucose-induced intestinal hyperemia is primarily characterized by premucosal A3 arteriole dilation in this model and that nitric oxide is a mediator of glucose-induced intestinal hyperemia. These findings suggest that either (1) glucose directly causes endothelial nitric oxide production or (2) epithelial cells transduce a vasodilatory signal through vascular endothelial-derived nitric oxide during postprandial intestinal hyperemia.

Analysis of Variance↗

PubMed1997

Detection of sperm within Gyrodactylus (Platyhelminthes, Monogenea) tissues using fluorescence and transmission electron microscopy.

A rapid fluorescent staining method demonstrating spermatozoa within gyrodactylid monogeneans is described. Gyrodactylids fixed and stained in 2% acetic acid containing 1 microgram ml-1 bisBenzimide (Hoechst 33258) were viewed using epifluorescence microscopy. In addition to staining sperm in the testis, seminal vesicle and seminal receptacle, the technique highlighted sperm in the gut and cytoplasmic lining of the uterus, locations from which they had not previously been recorded. The technique was more rapid than transmission electron microscopy (TEM), which was used to confirm the observations. Fluorescence microscopy provided an overview of sperm orientation and distribution that could otherwise be obtained only from serial ultrathin sections. Using the fluorescence technique and/or TEM we have located migrating sperm in the tissues of Gyrodactylus turnbulli, G. bullatraudis and G. gasterostei, although these sperm do not appear to enter the tissues of embryos in utero. The technique can be used to study insemination patterns in gyrodactylid populations and in experimental studies of gyrodactylid reproduction.

Animals↗