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Free-feeding patterns of rats: effects of pyrogen and dietary protein content.

Infusion of bacterial pyrogen (Priomen) was accompanied by an increase in body temperature, an increase in heat production, and a decrease in the voluntary food intake ofrats fed high-as well as low-protein diets. The magnitude of this pyrogen-induced depression of food intake was comparable for both diets. However, in rats fed high-protein diets, this decrease was additive to that normally seen following administration of such diets. These data indicate that the control of food intake cannot be explained in terms of a behavioral the more regulatory response.

Animals↗

Pyrogens fail to produce fever in a cordylid lizard.

We investigated the effects on body temperature of the lizard Cordylus cataphractus of intracardiac injections of leucocyte pyrogen (LP) synthesized from rabbit blood and of killed Aeromonas hydrophila, a gram-negative bacterium reputed to be pathogenic in lizards. Lizards were placed in a photothermal gradient that allowed them to select a preferred body temperature following the injections. Neither injection of 0.5 ml rabbit LP nor of 4 X 10(9) organisms of A. hydrophila in 0.2 ml sterile saline caused body temperature of lizards to differ from that of control lizards injected with sterile saline. Following injection of these solutions in the lizards placed in a thermal gradient where ambient temperature ranged from 20-88 degrees C, body temperature was maintained between 32 and 34 degrees C. Pyrogens failed to elevate body temperature even when body temperature was elevated artificially to 36 degrees C before injection. We conclude that C. cataphractus does not respond with fever to either rabbit LP or A. hydrophila. Fever may not be ubiquitous even among lizards.

Aeromonas↗

Pattern differences in experimental fevers induced by endotoxin, endogenous pyrogen, and prostaglandins.

To distinguish pattern differences in experimentally induced fevers, we investigated febrile responses induced by intravenous (IV), intracerebroventricular (ICV), and intra-preoptic/anterior hypothalamic (POA) administration of bacterial endotoxin (lipopolysaccharide, LPS), endogenous pyrogen (EP), human recombinant interleukin-1 alpha (IL-1), and prostaglandins E2 and F2 alpha (PGE2 and PGF2 alpha). Intravenous LPS, EP, or IL-1 in high concentrations caused biphasic fever. In low concentrations, they induced only the first phase of fever. Latency to onset and time to first peak of fever induced by IV injection of LPS or EP were almost the same as those after ICV or POA injection of PGE2. Fever induced by ICV or POA administration of LPS, EP, IL-1, or PGF2 alpha had a long latency to onset and a prolonged time course. There were significant differences among the latencies to fever onset exhibited by groups that received ICV or POA injections of LPS, EP, or PGF2 alpha and by groups given IV injections of LPS or EP and ICV or POA injections of PGE2. Present observations indicate different patterns of fever produced by several kinds of pyrogens when given by various routes. These results permit us to consider the possibility that there are several mediators or multiprocesses underlying the pathogenesis of fever.

Animals↗

Prostaglandin formation in the hypothalamus in vivo: effect of pyrogens.

Conscious cats were used to study the local release of prostaglandin (PG) E2 and thromboxane (Tx) B2 (the stable TxA2 by-product) from the preoptic-anterior hypothalamus (AH-POA) and the tuberal-posterior hypothalamus (PH-Tu) using a modified "push-pull" perfusion procedure. In the absence of fever, PGE2 release was steady from the 2nd h of perfusion onward, its rate at either site ranging between 0.08 and 0.12 pg/min. Local treatment with probenecid (1 mM) increased PGE2 release about threefold. Compared with PGE2, basal release of TxB2 was greater (0.15-0.43 pg/min) and, occasionally, tended to fall with time. Both compounds were found in higher amounts (2- to 10-fold increase) after locally injecting endotoxin, and the effect was greater in AH-POA than PH-Tu. Conversely, intravenous endotoxin (bolus) or interleukin 1 (IL-1) (bolus plus infusion) at doses causing a sustained fever selectively stimulated the formation of PGE2, but the response itself did not differ between AH-POA and PH-Tu. In either region, the degree of enhancement in PGE2 release correlated with the magnitude of the fever. Intravenous indomethacin (2 mg/kg) reversed both the fever and PGE2 elevation. These findings support an intermediary role for PGE2 in the central action of pyrogens and the ensuing fever. Blood-borne pyrogens may act at multiple sites in brain, which are tentatively identified with the circumventricular organs.

Animals↗

Altered interleukin-1 and tumor necrosis factor production and secretion during pyrogenic tolerance to LPS in rabbits.

Rabbits were injected intravenously with 10 micrograms/kg of endotoxin [lipopolysaccharide (LPS)] on days 0, 1, and 7, and rectal temperatures were monitored. The febrile responses were compared with circulating levels of interleukin-1 beta (IL-1 beta) and tumor necrosis factor (TNF) and in vitro synthesis of these cytokines by peripheral blood mononuclear cells (PBMC) isolated just before the injection of LPS. Fever after the first LPS injection was biphasic on day 0, attenuated and monophasic after the second LPS injection on day 1, and augmented after third injection of LPS on day 7. On day 1, circulating TNF and IL-1 beta levels were significantly (P < 0.05) decreased compared with those on days 0 and 7. Similarly, TNF and IL-1 beta synthesis by LPS-stimulated PBMC were significantly reduced on day 1. On day 7, cellular synthesis and secretion of IL-1 beta were significantly increased compared with that on day 0. A significant positive correlation was observed between fever index and total in vitro IL-1 beta synthesis by LPS-stimulated PBMC (r = 0.866, P = 0.001). These data demonstrate that pyrogenic tolerance in the rabbit after a single LPS injection is associated with decreased circulating IL-1 beta and TNF levels as well as decreased production of these cytokines in vitro. In addition, the pyrogenic hyperresponsiveness to LPS after 7 days is associated with increased synthesis and secretion of IL-1 beta from PBMC in vitro.

Animals↗

Fever suppression by subdiaphragmatic vagotomy in guinea pigs depends on the route of pyrogen administration.

It has recently been shown that subdiaphragmatic vagotomy blocks some of the effects of inflammatory stimuli on brain-controlled functions. Therefore, vagal afferent fibers have been proposed to play a prominent role in the communication pathways between the immune system and the brain. In the present study, we investigated the effect of subdiaphragmatic vagotomy on fever induced by intraperitoneal or intramuscular injection of lipopolysaccharide (LPS) or muramyl dipeptide (MDP), which are both cytokine-inducing agents in guinea pigs. Intraperitoneal and intramuscular injections of LPS or MDP were tested in the same animal with an interval of 1 wk. In one experiment, intraperitoneal injections of LPS or MDP were performed at the beginning, followed by intramuscular injections 1 wk later. In another experiment, intramuscular injections of LPS or MDP were performed at the beginning, followed by intraperitoneal injections 1 wk later. The febrile response to intraperitoneal injection of LPS was almost completely abrogated in vagotomized animals compared with sham-operated controls (from 30-330 min after intraperitoneal injection of LPS). The suppression of LPS fever was quantitatively the same in both experiments. In contrast, the response to intramuscular injection of LPS was the same in vagotomized and sham-operated guinea pigs in each of the experiments. Fever induced by intraperitoneal injection of MDP was partly attenuated by subdiaphragmatic vagotomy (between 150 and 300 min after intraperitoneal injection of MDP) in both experiments. Again, the febrile response to intramuscular injections of MDP was not significantly altered by subdiaphragmatic vagotomy. In conclusion, the suppressive effect of subdiaphragmatic vagotomy on fever induced by peripheral immune stimuli depends on the route of pyrogen administration. Because the febrile response to intraperitoneal injection of bacterial pyrogens is strongly diminished in vagotomized guinea pigs, it is suggested that vagal afferent fibers play a crucial role in the transduction of immune signals from the abdominal cavity to the brain.

Acetylmuramyl-Alanyl-Isoglutamine↗

Postnatal development of pyrogenic sensitivity in guinea pigs.

Despite evidence of thermoregulatory ability from birth, neonates generally are unable to develop fever when challenged with endotoxin. This could be due to their small capacity for heat storage. To test this possibility, the pyrogenicity of S. enteritidis endotoxin (2 mug/kg, iv) was measured at both room (Ta = 25 degrees C) and neutral (Tn = 29-33 degrees C, depending on age) temperatures in 0- to 32-day-old unanesthetized guinea pigs, reared from birth at about 24 degrees C. Control guinea pigs received sterile saline injections in concurrent experiments. Shivering, O2 uptake, and colonic (Tre) and subcutaneous [over the interscapular fat pad (Tbat) and the sacrospinalis muscle (Tsc)] temperatures were recorded continuously for 4 h after injection. Endotoxin generally produced no febrile responses at both ambient temperaturess rises in animals aged 8 or more days; Tbat increased before the other sites in the 8- and 16-day-old animals, and shivering did not occur; by 32 days of age, however, Tbat no longer increased first, and there was shivering. In Tn significant febrile rises were not evident until 32 days of age; control temperatures, however, were elevated during this exposure as compared to at Ta. These results showed therefore that pyrogenic sensitivity is not apparent in guinea pigs during the first postnatal week; thereafter fever responses are evocable, but their detection may be masked by environmentally produced changes in body temperature. The data also indicated that the site of the heat production underlying, in part, endotoxic fevers gradually shifts from brown fat so skeletal muscle during the first month of life.

Age Factors↗

Fever in the elderly. Production of leukocytic pyrogen by monocytes from elderly persons.

In an attempt to explain the diminished febrile response of the elderly, we studied the first step in fever generation, that of production of leukocytic pyrogen (LP) by monocytes. Monocytes from 25 healthy elderly volunteers (ages 65-91) and 24 healthy young volunteers (ages 17-38) were stimulated with Staphylococcus epidermidis to release LP; LP activity in the culture supernatants was assayed by measuring the pyrogenic response in rabbits and rats and the fall in plasma iron and zinc in rats. Monocytes from elderly volunteers produced slightly less LP than monocytes from young volunteers, but the difference was not statistically significant. The amount of LP produced was not correlated with age. Therefore, the diminished febrile response of the elderly is not the result of an intrinsic defect in the monocyte's ability to make LP. Other explanations relating to the central effect of LP and the effector response to LP in the elderly should be sought.

Adolescent↗

[Pyrogen-free sterile water for operative endoscopy (author's transl)].

In transurethral endoscopic operations we have maximal demands for the water conditions where, as in urology, for the diagnostic endoscopy one mostly uses sterile filtered but on principle hydrant water containing pyrogen. First the determinations of the order for drinking water and its possible consequences for irrigator systems are laid down. Water for injections like the irrigator water in operative endoscopy must conform to the demands in pharmacopoeia of the different countries. With the new combination of reverse osmosis system with an irrigator, hydrant water can be purified to pyrogen-free sterile water in the place of treatment.

Female↗

Human leukocytic pyrogen induces release of specific granule contents from human neutrophils.

The ability of highly purified human leukocytic pyrogen (LP) to induce neutrophil lysosomal protein release is described. Human peripheral blood neutrophils isolated by Ficoll-Hypaque and dextran sedimentation were exposed to purified human LP. The specific granule-associated proteins, lysozyme and lactoferrin were selectively released, whereas primary granule (beta-glucuronidase) and cytoplasmic (lactic dehydrogenase) enzyme markers were not. Optimum release was observed after 45 min in the presence of Ca++ and Mg++. Cytochalasin B (5 microgram/ml) had no effect on LP-induced lysosomal enzyme release. Since the pyrogenicity of LP is dependent on prostaglandin synthesis, the effect of two potent inhibitors of prostaglandin synthesis on lysozyme release was studied. Both indomethacin and naproxen failed to inhibit specific granule protein release. These observations suggest that the concommitance of fever, elevated serum or urine lysozyme and hypoferremia may, in part, be explained by the interaction of LP and peripheral blood neutrophils.

Cations, Divalent↗

[Measurement of endotoxin in blood products using an endotoxin-specific Limulus test reagent and its relation to pyrogenic activities in rabbit].

The amounts of endotoxin in commercial blood products were measured by the turbidimetric kinetic Limulus test with an ordinary reagent (LAL-HS) and a new endotoxin-specific reagent (LAL-ES). LAL-ES contains a sufficient amount of a water-soluble (1----3)-beta-D-glucan derivative as a blocker of the (1----3)-beta-D-glucan-mediated coagulation pathway in the reaction of the Limulus amebocyte lysate. The amounts of endotoxin in albumin and globulin products measured with LAL-ES agreed with pyrogenic activities in rabbits, but those measured with LAL-HS did not. Added endotoxin in the blood products was well recovered with LAL-ES, but that in some products was excessively recovered with LAL-HS. The amounts of endotoxin in diphtheria-pertussis-tetanus combined vaccines measured with LAL-HS and LAL-ES agreed with the pyrogenic activities in rabbits. The results suggested the existence of a false-positive substance like beta-glucan in the blood products but not in the vaccine. LAL-ES is more suitable for the detection of endotoxin in blood products than LAL-HS.

Animals↗

[Application of the limulus amebocyte lysate test to measurement of endotoxin in therapeutic human plasma protein fraction. Comparison with the rabbit pyrogen test].

We applied the limulus amebocyte lysate (LAL) test to the detection of bacterial endotoxins in therapeutic human plasma protein fraction (PPF) and compared the LAL-test with the rabbit pyrogen test. Two endotoxin-specific LAL-reagents were used for the colorimetric method and turbidimetric kinetic method. The amounts of added endotoxin to the PPF were correctly estimated by either method. The results of four independent assays for the 53 samples of PPF corresponded well with each other (correlation coefficient: 0.851-0.959, regression coefficient: 0.898-1.151). The amounts of endotoxin in the PPF estimated by the LAL-test significantly correlated with the rise of body temperature in rabbits (correlation coefficient: 0.547-0.642, and 0.911-0.934 for the endotoxin added samples). These results suggest that the LAL-test could be used as an alternative method for the rabbit pyrogen test to PPF.

Animals↗

Role of the nitric oxide/cyclic GMP/Ca2+ signaling pathway in the pyrogenic effect of interleukin-1beta.

Interleukin-1beta (IL-1beta) has a wide spectrum of inflammatory, metabolic, haemopoietic, and immunological properties. Because it produces fever when injected into animals and humans, it is considered an endogenous pyrogen. There is evidence to suggest that Ca2+ plays a critical role in the central mechanisms of thermoregulation, and in the intracellular signaling pathways controlling fever induced by IL-1beta and other pyrogens. Data from different labs indicate that Ca2+ and Na+ determine the temperature set point in the posterior hypothalamus (PH) of various mammals and that changes in Ca2+ and PGE2 concentrations in the cerebrospinal fluid (CSF) of these animals are associated with IL-1beta-induced fever. Antipyretic drugs such as acetylsalicylic acid, dexamethasone, and lipocortin 5-(204-212) peptide counteract IL-1beta-induced fever and abolish changes in Ca2+ and PGE2 concentrations in CSF. In vitro studies have established that activation of the nitric oxide (NO)/cyclic GMP (cGMP) pathway is part of the signaling cascade transducing Ca2+ mobilization in response to IL-1beta and that the ryanodine (RY)- and inositol-(1,4,5)-trisphosphate (IP3)-sensitive pools are the main source of the mobilized Ca2+. It is concluded that the NO/cGMP/Ca2+ pathway is part of the signaling cascade subserving some of the multiple functions of IL-1beta.

Animals↗

Direct effects of endogenous pyrogen on medullary temperature-responsive neurons in rabbits.

The effect of endogenous pyrogen (E.P.) injected directly into the tissue near the recording site were examined on the activities of the medullary temperature-responsive (TR) neurons in rabbits anesthetized with urethane. Endogenous pyrogen prepared from rabbit's whole blood was administered by a fine glass cannula (100-200 micrometer in diameter) in a fluid volume of 1 to 4 microliter. The cannula was fixed to the manipulator in parallel with a microelectrode and their tips were less than 0.05 mm apart. In rabbits with the intact preoptic/anterior hypothalamic (PO/AH) region, 4 warm-responsive neurons out of 7 were inhibited and 6 cold-responsive neuron out of 7 were excited by the direct administration of the E.P. In rabbits with lesions of the PO/AH, 5 warm-responsive neurons out of 9 were inhibited and 6 cold-responsive neurons out of 8 were facilitated by E.P. Antipyretics administered locally after the E.P. antagonized the pyretic effect, causing a return of the discharge of TR neuron to the control rate within 2.4 +/- 1.2 (mean +/- S.D.) min. The medullary TR neuron itself has the ability to respond to the E.P. and contributes to the development of fever.

Animals↗

Pyrogenicity of etiocholanolone and interleukin-1 in New and Old World Monkeys.

Etiocholanolone (5beta-androstan-3alpha-ol-17-one; designated E) is one of the major products of metabolism of testosterone and androstenedione (androst-4-ene-3,17-dione) in many mammalian species, including humans. E and several other 5beta-reduced steroids have been found to induce fever in humans. The pyrogenic effect of these steroids has been shown to be due to the release of interleukin-1 (IL-1) from the leukocytes that are mobilized in response to the steroid injections. Old World Monkeys such as Rhesus monkeys (Macaca mu/atta), metabolize androgens similarly to humans, and E is a normal metabolite. However, New World Monkeys such as Squirrel monkeys (Saimiri sciureus), lack hepatic 5alpha- and 5beta-steroid reductases and excrete androgens primarily in an unaltered state; E is not produced. Therefore, we postulate that Squirrel monkeys likewise may have lost the ability to respond to 17-ketosteroids such as E. To test this hypothesis, adult male Rhesus and Squirrel monkeys were treated with E, and their rectal temperatures were recorded over a 24-hr period. Rhesus monkeys exhibited a rise of up to 3 degrees F following E injection. Squirrel monkeys, on the other hand, did not exhibit any increase in rectal temperature over the 24-hr period, even when doses up to 250 times the effective human dose were used. However, both species responded to injected IL-1alpha with a robust increase in rectal temperature. The data show that E is pyrogenic in Rhesus, but not Squirrel monkeys. The findings support the notion that injected E may induce release of IL-1 in Rhesus monkeys, but not in Squirrel monkeys.

Androsterone↗

Development of streptococcal pyrogenic exotoxin C vaccine toxoids that are protective in the rabbit model of toxic shock syndrome.

Streptococcal pyrogenic exotoxin C (SPE C) is a superantigen produced by many strains of Streptococcus pyogenes that (along with streptococcal pyrogenic exotoxin A) is highly associated with streptococcal toxic shock syndrome (STSS) and other invasive streptococcal diseases. Based on the three-dimensional structure of SPE C, solvent-exposed residues predicted to be important for binding to the TCR or the MHC class II molecule, or important for dimerization, were generated. Based on decreased mitogenic activity of various single-site mutants, the double-site mutant Y15A/N38D and the triple-site mutant Y15A/H35A/N38D were constructed and analyzed for superantigenicity, toxicity (lethality), immunogenicity, and the ability to protect against wild-type SPE C-induced STSS. The Y15A/N38D and Y15A/H35A/N38D mutants were nonmitogenic for rabbit splenocytes and human PBMCs and nonlethal in two rabbit models of STSS, yet both mutants were highly immunogenic. Animals vaccinated with the Y15A/N38D or Y15A/H35A/N38D toxoids were protected from challenge with wild-type SPE C. Collectively, these data indicate that the Y15A/N38D and Y15A/H35A/N38D mutants may be useful as toxoid vaccine candidates.

Animals↗

[Detection of pyrogenic exotoxin SpeA, SpeB and SpeC genes in Chilean streptococci isolates and their association with clinical manifestations].

BACKGROUND: The virulence of Streptococcus pyogenes is determined by a variety of structural molecules, toxins and complex enzymes. Pyrogenic exotoxins cause fever, erythematous reactions, cytotoxic and immunological effects. AIM: To assess the frequency of speA, SpeB and SpeC genes in Chilean Streptococcus pyogenes strains and their association with the invasiveness of infections. MATERIAL AND METHODS: The genes for pyrogenic exotoxins SpeA, SpeB and SpeC were determined by polymerase chain reactions in 114 strains of group A Streptococcus pyogenes isolated from Chilean patients with invasive or non invasive infections. RESULTS: The gene for SpeA was present in 30.7% of isolates, the gene for SpeB was present in 69.3% and the gen for SpeC in 44.7% of isolates. The gene for SpeA was present in 20 of 33 invasive infections and in 15 of 81 non invasive infections (p < 0.0001). On the contrary, the gene for SpeC was present in 11 of 33 invasive infections and in 41 of 81 non invasive infections (p < 0.05). The frequency of speB was similar in invasive and non invasive infections. CONCLUSIONS: There is a clear relationship between the presence of SpeA genes and the severity of infections caused by Streptococcus pyogenes.

Bacterial Proteins↗

[The endotoxin from S-, R- and M-forms of enteropathogenic E. coli O 149: pyrogenicity, Shwartzman-phenomenon and hypersensitivity (author's transl)].

The authors are interested in elucidation of the role of endotoxin from S, R- and M-forms of E. coli O 149, and of the role of hypersensitivity in pathogenesis of Colibacillosis. It was shown in this experiment that all S, R- and M-form of E. coli O149 possess endotoxin with almost the same pyrogenicity in normal rabbits. The pyrogenicity increases enormously in the hypersensitized rabbit. There were also not observed differences between endotoxins extracted from S, R, M-mutnants. All 3 endotoxins evoke in normal rabbits the skin Shwartzman-Phenomenon, the S-endotoxin was there much stronger. This phenomenon increases in capacity in hypersensitized rabbits. Thus again, the role of hypersensitivity in Colibacillosis was proved to be important.

Animals↗