Biomedical subjects

P D Harris

Publications and source records attributed to P D Harris.

At least 19 recordsLinked to original sources

PubMed2003

Degradation of extracellular matrix components by defined proteinases from the greenbottle larva Lucilia sericata used for the clinical debridement of non-healing wounds.

BACKGROUND: Larvae of the greenbottle fly Lucilia sericata are used routinely for the clinical treatment of difficult necrotic and infected wounds. Degradation by proteinases contained in larval excretory/secretory (ES) products is thought to contribute to wound debridement by removal of dead tissue. However, proteinase activity may also affect host tissue remodelling processes. OBJECTIVES: To identify proteolytic enzymes derived from L. sericata ES products with activities against fibrin and extracellular matrix (ECM) components. METHODS: Larval proteinase activities were assayed in vitro using class-specific substrates and inhibitors. Their action against fibrin and ECM components was examined using sodium dodecyl sulphate-polyacrylamide gel electrophoresis. RESULTS: Three classes of proteolytic enzyme were detected in the secretions using fluorescein isothiocyanate-labelled casein as a model substrate. The predominant activity belonged to serine proteinases (pH optima 8-9) of two different subclasses (trypsin-like and chymotrypsin-like), with a weaker aspartyl proteinase (pH 5) and a metalloproteinase (pH 9) with exopeptidase characteristics also present. Using skin-relevant ECM components as substrates L. sericata ES products solubilized fibrin clots and degraded fibronectin, laminin and acid-solubilized collagen types I and III. Hydrolysis of ECM macromolecules was inhibited by preincubating ES products with phenylmethylsulphonyl fluoride but not 4-amidinophenylmethylsulphonyl fluoride, indicating that degradation was due to the 'chymotrypsin-like' serine proteinase. CONCLUSIONS: These data suggest that a combination of L. sericata ES proteinases involving chymotrypsin-like and trypsin-like activities could potentially influence wound healing events when maggots are introduced into necrotic and infected wounds, with the chymotrypsin-like activity involved in the remodelling of ECM components.

Animals↗

PubMed2002

Gyrodactylid developmental biology: historical review, current status and future trends.

In the viviparous gyrodactylids, embryos develop one inside another within the parental uterus, a phenomenon with major implications for the biology of this species-rich group. Development occurs via two routes: first-born daughters develop at the centre of an embryo cluster in utero, whereas all other daughters develop from oocytes. The resulting offspring are, however, morphologically indistinguishable. We review here the history of gyrodactylid embryology in the context of current knowledge and, present additional cytogenetic and ultrastructural observations of embryonic development. These progenetic parasites are highly modified for viviparity; oocyte maturation and sperm storage occur in a single chamber, the Egg Cell Forming Region, and a mature oocyte passes into the uterus after the birth of the preceding, fully developed offspring. The uterus has a syncytial lining derived from anterior and posterior cap cells. These cells are the first to differentiate in the female reproductive system and may be involved in controlling development. Embryos receive nutrients via the uterus rather than from vitelline cells. Traditionally, development of the first-born daughter has been considered a form of polyembryony, although paedogenesis has also been suggested. In contrast to previous studies, we could not trace lineage of the first-born daughter to a single quiescent macromere. However, only mitotic divisions have been conclusively observed in the intraembryonic generation, indicating an asexual origin. All other daughters are formed from meiotically derived oocytes by sexual reproduction or automictic parthenogenesis. The latter may involve pre-meiotic doubling of chromosomes, but the precise mechanism and the relative proportion of sexual and parthenogenetic offspring are unknown. Exceptionally, cleavage in Gyrodactylus spp. occurs by duets rather than quartets (a pattern previously only recorded in acoels) and is characterised by extensive cell rearrangements. Blastomeres may be connected by fine cytoplasmic processes or completely disassociated and are readily redistributed by the muscular actions of the parental uterus. This process resembles 'Blastomeren-Anarchie' of rhabdocoels but without the structural support of vitelline cells. It prevents generation of early cell fate maps and indicates regulative, rather than mosaic, development. Structures such as the gut and excretory system differentiate late, and are highlighted, together with the attachment apparatus, as examples of post-embryonic differentiation. Molecular and cellular techniques are now essential to further elucidate mechanisms of gyrodactylid reproduction, which will in turn contribute to current debates with animal embryology.

Animals↗

PubMed2002

Endogenous norepinephrine regulates blood flow to the intact rat tibia.

The goal of our study was to determine if endogenous norepinephrine (NE) has a role in the regulation of basal blood flow to intact bone. The experimental plan was to measure bone blood flow before and after pharmacological blockade of alpha-adrenergic receptors. A significant increase in blood flow after receptor blockade would suggest that endogenous norepinephrine exerts a tonic constrictor effect on the vessels supplying blood to the bone. Mature, male rats were anesthetized with Inactin. Arterial blood pressure and left tibia blood flow (laser Doppler flowmetry) were measured. A cannula was inserted into the right iliac artery and advanced to the aortic bifurcation to deliver drugs into the left hindlimb circulation, including the left tibia vasculature. Bolus injection of norepinephrine caused a dose-dependent decrease in bone blood flow (30-40%). Blockade of alpha-adrenergic receptors with phentolamine or phenoxybenzamine attenuated by more than 50% the norepinephrine-induced decrease in bone blood flow. In separate rats that had not received exogenous norepinephrine, injection of phentolamine alone decreased bone vascular resistance by 34+/-3%. Similarly, phenoxybenzamine decreased resistance by 25+/-4%. These results are consistent with the conclusion that alpha-adrenergic receptors mediate a significant constriction of blood vessels which participate in the partial control of basal blood flow to the intact rat tibia.

Adrenergic alpha-Antagonists↗

PubMed2002

Survival, feeding and embryo development of Gyrodactylus gasterostei (Monogenea: Gyrodactylidae).

Viviparous gyrodactylid monogeneans which are detached from their host may form an important source of infection in fish communities. This is particularly significant for these ectoparasites which have no specific free-living stage and where transmission usually occurs through transfer of adult parasites when fish come into close contact. In this study, in vitro survival of Gyrodactylus gasterostei was correlated with changes in tissue ultrastructure of parents and their embryos during detachment and following return to a host. At 10 degrees C, detached worms survived for up to 89 h (50%) survival = 20 h) with mortality influenced by both age-independent and age-dependent factors. The gastrodermis of attached, fed parasites is syncytial and contains a variety of vesicles and feeding vacuoles. However, following detachment and starvation, degenerative changes occur within the gastrodermis, eventually leading to marked damage to the embryo/s. When starved worms reattached to a fish, they began browsing on host epithelial cells within 1 min and ultrastructural evidence for phagocytic activity in the gastrodermis was detected after 5 min. Putative waste vacuoles increased in abundance after 5-30 min on the new host, coinciding with the reappearance of electron-dense vesicles and possibly indicating completion of the first intracellular cycle of digestion. Parental feeding directly influenced the normal cyclical maturation and regression of the uterine lining between births. In extreme cases, starvation led to detached parasites (6.1%) aborting their embryos. However, even short periods off the host influenced development and survival of embryos, suggesting that temporary interruption of nutrient flow to the embryo can significantly affect gyrodactylid reproductive rates.

Animals↗

PubMed2002

Behavior favoring transmission in the viviparous monogenean Gyrodactylus turnbulli.

Transmission by Gyrodactylus turnbulli occurs most frequently when its hosts (Poecilia reticulata) come into close contact. This study is the first description of a specific migratory behavior that facilitates transmission of a gyrodactylid from dead hosts. Recently-dead guppies typically float at the water's surface; G. turnbulli moves off these fish into the water film, hanging motionless with the haptor held by surface tension. Because guppies are surface feeders, detached parasites in the water film are more likely to contact a new host.

Animals↗

PubMed2001

Sepsis increases NOS-2 activity and decreases non-NOS-mediated acetylcholine-induced dilation in rat aorta.

INTRODUCTION: Acetylcholine (Ach) is frequently used to assess endothelium-dependent vasodilation during sepsis. However, the effects of sepsis on constitutive nitric oxide synthase activity (NOS-1 and -3) and other non-NOS effects of Ach are unclear. METHODS: Sepsis was induced in rats by inoculation of an implanted sponge with Escherichia coli and Bacteroides fragilis (10(9) CFU each). Thoracic aortic rings (2 mm) were harvested at 24 h from septic (N = 9) and control (N = 9) rats and were suspended in physiological salt solution (PSS), PSS + l-N(6)-(1-iminoethyl)lysine (l-NIL: NOS-2 inhibitor, 10 microM), or PSS + l-N(G)-monomethylarginine (l-NMMA: NOS-1, -2, and -3 inhibitor, 60 microM). Rings were set at 1-g preload and precontracted with phenlyephrine (10(-8) M). Relaxation dose-response curves were generated with six doses of Ach (3 x 10(-8) to 10(-5) M). RESULTS: Sepsis increased the maximal relaxation to Ach under basal conditions. NOS 2 inhibition with l-NIL decreased Ach-induced relaxation in controls (66% vs 84%, P < 0.05, two-way ANOVA) and more so in septic rats (44% vs 93%, P < 0.05). Total NOS inhibition with l-NMMA decreased Ach-induced relaxation to 45% (P < 0.05) in controls and to 30% (P < 0.05) in septic animals. CONCLUSIONS: Inhibition of NOS-1, -2, and -3 failed to abolish Ach-induced relaxation, suggesting the presence of other Ach-induced vasodilator mechanisms. NOS-2 inhibition reduced Ach-induced relaxation by 20-25% in the normal thoracic aorta, but by 50% in septic animals. The remaining Ach-induced non-NOS vasodilation (after inhibition of NOS-1 + NOS-2 + NOS-3) was reduced from 45% in normals to 30% in septic animals. Vascular dysregulation in sepsis is a complex event involving increased NOS-2, decreased NOS-1 + NOS-3, and decreased Ach-induced non-NOS vasodilator mechanisms.

Acetylcholine↗

PubMed2001

Progressive decrease in constrictor reactivity of the non-absorbing intestine during chronic sepsis.

Chronic sepsis leads to an impaired intestinal microcirculation, which might reflect altered microvascular control. We hypothesized that intestinal microvascular sensitivity to norepinephrine (NE) is decreased during chronic sepsis. Chronic sepsis was induced by a polymicrobial inoculation of implanted subcutaneous sponges in rats. Septic rats were studied either 24 or 72 h after a single inoculation (1-hit) of bacteria. Other rats received a second inoculation (2-hit) of bacteria 48 h later and were studied at 24 h after the second inoculation. NE (0.01-1.0 microM) responses in the non-absorbing terminal ileal arterioles (inflow A1, proximal-p and distal-d premucosal A3) were measured by video microscopy. NE threshold sensitivity (pD(T20) = -log of 20% response dose) was analyzed. pD(T20) was significantly decreased in A1, pA3, and dA3 of 1-hit 24-h septic rats (P < 0.05), and was further decreased in all vessels of 2-hit 72-h septic rats (P < 0.05). In contrast, the pDT(T20) of all three vessels significantly returned toward normal values after 72 h in rats that had only 1 bacteria inoculation. We conclude that an initial bacterial challenge decreases vasoconstrictor reactivity of the intestinal microcirculation and that subsequent repeated bacterial challenge exacerbates this defect in vasoconstrictor control in the non-absorbing intestine.

Animals↗

PubMed2001

Ultrasonic transmission and reception from bulk-micromachined transducers.

Acoustic transduction in air from two bulk-micromachined silicon structures is investigated. Both contain silicon diaphragms of the order of 2 mm2 in close proximity to a metallized substrate. One diaphragm is mass-loaded; the other is not. Their resonant frequencies (70 and 360 kHz) are dominated by squeeze film trapping of ambient air, and the Q of each device is about 8. The lower frequency (LF) device is characterized by electrical and acoustic measurements using a calibrated microphone. Novel diagnostic methods that exploit the non-linear nature of the transducer are described. The adequacy of calibration by reciprocity is confirmed at 70 kHz and applied to the high frequency device. An insertion loss of 19 dB is measured, which compares well with reports of other silicon-based transducers. Observed losses are accounted for by squeeze-film damping applied to the diaphragm-substrate gap. The ability to control the bandwidth by the squeeze film effect, the good efficiency, and the relatively standard method of construction could make such ultrasonic transducers useful in specialist applications.

Journal Article↗

PubMed2000

Population growth of Gyrodactylus salaris (Monogenea) on Norwegian and Baltic Atlantic salmon (Salmo salar) stocks.

Reproductive success of Gvrodactylus salaris from River Lierelva was compared experimentally on 3 stocks of salmon (12.5 degrees C +/- 0.2). Isolated fish from 2 susceptible Norwegian stocks (Rivers Lier and Alta) and 1 resistant Baltic stock of salmon (River Neva) were infected with a single gravid worm in order to record the temporal sequence of births and age at death of individual parasites. Establishment success (proportion of worms surviving to give birth) was generally low and mortality high, but significantly fewer worms survived on Neva (45% with mean survival of 3.5 days) compared to Alta and Lier fish (60 % mean survival 7.9 and 5.2 days, respectively). There was a dramatic difference in parasite fecundity between the host stocks: only 2 births occurred on Neva fish compared to third and fourth births on both Alta and Lier hosts. The timing of the first birth was more variable on Neva hosts and was significantly extended (mean 2.3 days) relative to that on Alta and Neva fish (1.8 days). However, timing of the second birth did not vary on any of the 3 salmon stocks. Age-specific mortality and fecundity data are consistent with exponential population growth of G. salaris on Alta and Lier fish but eventual extinction on Neva hosts. This is the first demonstration that gyrodactylids maintained on different host stocks exhibit variations in fecundity, development and mortality, which may in turn account for the variable virulence noted when G. salaris infects different salmonid hosts.

Age Factors↗

PubMed2000

Increased susceptibility of salmonids to the monogenean Gyrodactylus salaris following administration of hydrocortisone acetate.

Gyrodactylus salaris infects numerous salmonid species, ranging from the fully susceptible (Norwegian strains of Salmo salar), through species which, though initially susceptible, eventually eliminate their infections (Salvelinus alpinus and S. fontinalis) to entirely resistant (Salmo trutta) species. Here we describe experiments in which Salvelinus alpinus, S. fontinalis and Salmo trutta, implanted with hydrocortisone acetate to simulate stress-induced immunosuppression, were challenged with G. salaris. With previously uninfected Salvelinus fontinalis, G. salaris infections on fish treated with hydrocortisone acetate grew larger, and for longer, than on sham-treated controls. A similar result was obtained with S. trutta. Patterns of infection on Arctic charr, Salvelinus alpinus, were more complex, because individual fish varied from susceptible to highly resistant. Fish were therefore initially infected with G. salaris, and the most highly resistant group of individuals identified and disinfected. After 6 months recovery from this primary infection, hydrocortisone acetate was administered to half the fish, and all were challenged with G. salaris. Parasite populations on the hydrocortisone-treated individuals were consistently larger than those on the sham-treated controls, exceeding 30 parasites per fish after 5 weeks, in comparison with less than 10 parasites per fish on controls. These results indicate that hydrocortisone administration can lead to enhanced gyrodactylid populations on a range of salmonids. This suggests that the response to G. salaris is mediated by the immune system, and that the spectrum of responses observed in different species are, at least in part, due to the same mechanism. At a practical level, stress-induced immunosuppression during handling and transport of cultured salmonids may prove an important factor in the dissemination of G. salaris between watersheds.

Animals↗

PubMed2000

Gyrodactylus poeciliae n. sp. and G. milleri n. sp. (Monogenea: Gyrodactylidae) from Poecilia caucana (Steindachner) in Venezuela.

Two new species of Gyrodactylus are described from the South American live-bearing fish Poecilia caucana from Venezuela. Gyrodactylus poeciliae n. sp. is characterised by conspicuous ventral bar processes and by marginal hooks in which the tips are barely curved. Morphologically, the species resembles G. bullatarudis and G. costaricensis, and its relationship to the former was confirmed by sequencing the Internal Transcribed Spacers (ITS-1 and ITS-2) and the 5.8S ribosomal gene of the ribosomal DNA. Gyrodactylus milleri n. sp. was characterised by large, sharply angled marginal hook sickles in which the tips overhang the toes. This species resembled most closely G. turnbulli from guppies. These descriptions confirm the occurrence of G. turnbulli and G. bullatarudis-like gyrodactylids on a range of South American poeciliid fishes.

Animals↗

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↗