Intramuscular Injection Of Turpentine Research Articles

Comparison of 2-amino-[3- 11C]isobutyric acid and 2-deoxy-2-[18F]fluoro-D-glucose in nude mice with xenografted tumors and acute inflammation

2-Deoxy-2-[18F]fluoro-D-glucose ([18F]FDG) accumulates in tumors and also during active inflammation, including therapy-related inflammation. Additional PET tracers that are less avid to inflammation could be useful in differentiating cancer from inflammation and could complement the limitation of [18F]FDG-PET. 2-Amino-[3-11C]isobutyric acid ([3-11C]AIB) is a potential PET tracer for this purpose. We compared [3-11C]AIB and [18F]FDG uptakes in tumors and acute inflammation in a mouse model. Acute inflammatory lesions were induced in the hind legs of tumor-bearing mice by intramuscular injection of turpentine, and we conducted biodistribution and dynamic PET studies on [3-11C]AIB and [18F]FDG. [3-11C]AIB tumor uptake increased with time and was statistically significantly higher than [18F]FDG uptake. In inflamed muscles, [3-11C]AIB uptake was statistically significantly lower than [18F]FDG uptake, and the tumor-to-inflammation ratio for [3-11C]AIB was statistically significantly higher than that for [18F]FDG. [3-11C]AIB accumulates more selectively in tumor tissue than does [18F]FDG and thus has the potential of discriminating between tumors and inflammatory lesions better and of complementing the limitation of [18F]FDG.

Detecting inflammation with 131I-labeled ornidazole

The aim of this study was to demonstrate the accumulation of 131I-labeled ornidazole ( 131I-ORN) in experimental abscesses. 131I-ORN was prepared by electrophilic radioiodination of ORN, using radioiodide in the presence of Iodogen®. An in vivo inflammation model was prepared by intramuscular injection of turpentine into the thigh of rabbits. Four days later 131I-ORN was intravenously administered to rabbits. Serial scintigrams were performed at different periods, using a Sophy DX Gamma Camera. 131I-ORN was visualized at 10 min after injection. 131I-ORN was also administered intraperitoneally to rats with turpentine-induced inflammation, for quantitative biodistribution studies. Counts of selected tissues were taken by a NaI(Tl) scintillation detector (gamma counter) after rats were decapitated. The target-to-non-target muscle ratios were 2.5, 2.6, 2.9 and 1.9 at 1, 3, 5 and 24 h, respectively.

Inhibition of Tumor Necrosis Factor-α Action within the CNS Markedly Reduces the Plasma Adrenocorticotropin Response to Peripheral Local Inflammation in Rats

The present study tested the hypothesis that the cytokine tumor necrosis factor-alpha (TNF-alpha) is an important CNS mediator of the hypothalamo-pituitary-adrenal (HPA) axis response to local inflammation in the rat. Recombinant murine TNF-alpha administered directly into the cerebroventricles of normal rats produced a dose-dependent increase in plasma adrenocorticotropin (ACTH) concentration. Local inflammation induced by the intramuscular injection of turpentine (50 microl/100 gm body weight) also produced an increase in plasma ACTH, peaking at 160-200 pg/ml at 7.5 hr after injection (compared with 10-30 pg/ml in controls). Intracerebroventricular pretreatment with either 5 microl of anti-TNF-alpha antiserum or 1-50 microg of soluble TNF receptor construct (rhTNFR:Fc) reduced the peak of the ACTH response to local inflammation by 62-72%. In contrast, intravenous treatment with the same doses of anti-TNF-alpha or rhTNFR:Fc had no significant effect on the ACTH response to local inflammation. Although these data indicated an action of TNF-alpha specifically within the brain, no increase in brain TNF-alpha protein (measured by bioassay) or mRNA (assessed using either in situ hybridization histochemical or semi-quantitative RT-PCR procedures) was demonstrable during the onset or peak of HPA activation produced by local inflammation. Furthermore, increased passage of TNF-alpha from blood to brain seems unlikely, because inflammation did not affect plasma TNF-alpha biological activity. Collectively these data demonstrate that TNF-alpha action within the brain is critical to the elaboration of the HPA axis response to local inflammation in the rat, but they indicate that increases in cerebral TNF-alpha synthesis are not a necessary accompaniment.

Accumulation of 99mTc-Cysteine in Experimental Inflammatory Lesions in Comparison to 99mTc-Glutamine and 99mTc-HIG

The aim of this experimental work was to investigate the efficacy of 99mTc-L-Cysteine (Cys) in scintigraphic visualization of inflammatory lesions in comparison to 99mTc-L-glutamine (G), and 99mTc-HIG. In mice abscesses were induced by intramuscular injection of turpentine. Six days later mice were injected with 3.7 MBq of each agent and sacrificed in groups of three at 1, 3, 6 and 24 h. Scintigrams were obtained with a gamma camera. The organs, some blood, abscesses, some muscle and urine were removed, weighed and counted in a gamma counter. Percentage of uptake by organs and per gram tissues and abscess/normal tissue concentration ratios were calculated. Experimental arthritis was produced in 6 New Zealand rabbits by intraarticular injection of ovalbumin. Four days later 37 MBq of 99mTc-Cys and 99mTc-HIG were each i.v. administered to 3 rabbits. Scintigrams obtained at 1, 3, 6, and 24 h demonstrated the arthritic joints very well. ROI's over arthritic joints were compared to contralateral normal joints (A/C). In mice the abscesses were well visualized on all scintigrams. The maximum abscess/muscle ratios were 5.21 +/- 1.09 (6 h), 3.73 +/- 0.81 (3 h) and 5.98 +/- 1.17 (24 h) and the maximum abscess/blood ratios were 3.46 +/- 1.33 (24 h), 1.81 +/- 0.10 (6 h) and 0.914 +/- 0.351 (24 h) for 99mTc-Cys, 99mTc-G, and 99mTc-HIG, respectively. In rabbits the maximum A/C ratios were 2.61 +/- 0.53 (3 h) and 2.92 +/- 0.99 (24 h) for 99mTc-Cys and 99mTc-HIG, respectively. Our results indicate that 99mTc-Cys is a promising agent for imaging inflammatory lesions. It is preferred to 99mTc-HIG, because of higher concentration ratios attained earlier, lower blood background, lower cost and a simpler in-house preparation method.

Polyacrylamide gel electrophoretic serum protein patterns of acute inflammation induced by intramuscular injection of turpentine in young broiler chickens.

Changes in the serum protein patterns accompanying inflammation in young broilers were examined by polyacrylamide gradient gel electrophoresis. Sera were obtained from blood of 4-week-old male broilers after induction of acute inflammation by an injection of 0.5 ml/kg turpentine. Electrophoretic patterns at 48 hr after injection, showed an increase in transferrin (Tf) and a segment containing albumin (Alb) with multiple peaks. The unbound iron binding capacity (UIBC) increased by 10 times, and the serum iron (SI) concentration in acute inflammation was reduced to one-half its initial value. These are considered to be typical changes in serum proteins and iron after acute inflammation.

Polyacrylamide Gel Electrophoretic Patterns of Chicken Serum in Acute Inflammation Induced by Intramuscular Injection of Turpentine

To develop a method to detect hidden inflammation using serum protein in chickens, changes in serum proteins with acute inflammation were analyzed using a turpentine-induced inflammation model. Inflammation in the pectoral muscle of a 14-wk-old White Leghorn became apparent 3 h after the injection of turpentine and became more severe thereafter. Coincident with the development of inflammation, changes in serum proteins were analyzed by electrophoresis on polyacrylamide gradient gels. The electrophoretic patterns were divided into 21 segments. Two of these segments increased remarkably. These were located near the center of the electrophoretic pattern and were identified as transferrin due to iron staining, correlation of movement against a commercial transferrin sample in SDS-PAGE, and immunoblotting. These results suggest that transferrin may serve as a marker for inflammation in chicken.

Tumor necrosis factor-alpha and fever after peripheral inflammation in the rat

The involvement of endogenous tumor necrosis factor-alpha (TNF-alpha) in the pyrogenic [i.e., rise in colonic temperature (Tc)] and thermogenic [increase in oxygen consumption (VO2)] responses to inflammation was investigated in rats subjected to an intramuscular injection of turpentine. Turpentine administration caused a rise in Tc and VO2 within 2 h (0.9 +/- 0.1 degrees C, 27 +/- 2%, respectively). Eighteen to twenty hours after turpentine, the magnitude of these responses had increased (2.3 degrees C fever and a 28% increase in metabolic rate compared with control animals) and was associated with marked inflammation in the injected limb. A rapid (by 4 h) and sustained rise in the plasma concentration of the endogenous pyrogen IL-6, but not TNF-alpha, was also observed. Intravenous pretreatment with a TNF-alpha antiserum attenuated the rise in Tc observed 2, 8, and 18 h after turpentine injection and almost abolished the hypermetabolic response observed at 18 h. In addition, the TNF-alpha antiserum inhibited the peak rise (8 h) in plasma IL-6 by 76%. These findings indicate that endogenous TNF-alpha is involved in fever and hypermetabolism during inflammation and that it may exert these effects by inducing the release of IL-6 into circulation.

The in-vitro uptake of zinc by blood cells in rats with long-term inflammatory stress

The aim of the present study was to investigate the in vitro uptake of zinc by blood cells, in an attempt to distinguish between those conditions in which low plasma zinc concentrations are due to inflammatory stress, and those which are due to true zinc deficiency. Inflammation induced by intramuscular injection of turpentine caused a significant reduction in plasma albumin concentrations, which persisted until the end of the study (2 weeks). It also caused a reduction in the plasma zinc concentration which was most marked during the first few days. A smaller difference persisted until the end of the study. When the serum zinc concentration was corrected for the hypoalbuminaemia, the changes in serum zinc concentration after the first 4 days of turpentine were small and mainly non-significant. The in vitro uptake of zinc by erythrocytes obtained from animals with inflammation did not increase, whereas the uptake was increased in cells obtained from animals with true zinc deficiency. Therefore this study suggests a method that can probably differentiate between an apparent zinc deficiency due to inflammatory stress and a real zinc deficiency, but additional experiments to validate this method should be performed.

Dietary n‐3 Fatty Acids Inhibit Fever Induced by Inflammation in the Rat

Modification of endogenous eicosanoid synthesis by dietary n-3 fatty acid supplementation reduces febrile responses, but the mechanisms underlying these effects in vivo have not been determined. In the present study, local inflammation was induced by intramuscular injection ofturpentine in rats fed control or n-3 supplemented diets for 8-9 weeks. In animals fed the control diet, turpentine induced fever, hypermetabolism, marked local inflammation (oedema), increased plasma IL-6 concentrations and raised cerebrospinal fluid (CSF) concentrations of PGE2. N-3 fatty acid supplementation significantly inhibited the rise in CSF PGE2, fever and hypermetaboHsm induced by turpentine. Local inflammation and increased plasma IL-6 concentrations were not affected by n-3 supplementation. These findings suggest that modification of dietary fat intake inhibits fever via reduced release of prostaglandins, probably within the brain, but does not affect the local or afferent signals involved in fever generation.

Mechanisms of early and late hypermetabolism and fever after localized tissue injury in rats

Inflammation and localized tissue injury were induced in rats by intramuscular injection of turpentine. This caused rapid and sustained (2-24 h) increases in colonic temperature (Tc; up by 2.5 degrees C) and oxygen consumption (VO2; up by 30%) as well as hypophagia (57%). The early responses (0-2 h) were attenuated by anesthesia, C-fiber deafferentation, peripheral but not central injection of a cyclooxygenase inhibitor, or central injection of a neutralizing antibody to corticotropin-releasing factor (CRF). In contrast, the later (18-20 h) changes in Tc and VO2 were unaffected by anesthesia, C-fiber deafferentation, or central injection of a CRF antibody but were inhibited by either peripheral or central administration of a cyclooxygenase inhibitor. beta-Adrenoceptor blockade reduced both phases of the response, and brown adipose tissue activity was significantly elevated from 4 to 24 h after turpentine injection. These data suggest that different mechanisms mediate the metabolic response to localized tissue injury, the early phase involving neural mechanisms characteristic of stress and inflammation, whereas the later phase appears to depend on humoral factors that are commonly associated with infection.

Acute Phase Proteins in Neonatal Rabbits

Acute phase protein response accompanies tissue injury, inflammation, or infection. During the acute phase, serum levels of C-reactive protein (CRP) can increase as much as 1,000-fold. We found that in response to an intramuscular injection of turpentine, neonatal rabbit CRP-specific RNA and serum CRP rose minimally. In contrast, adult serum levels of CRP increased 20-fold and mRNA for CRP in adults increased commensurately. However, during neonatal acute phase reactions, changes in the synthesis of the third component of complement (C3) and albumin were induced, implying a dysynchronous development of the response of various acute phase proteins.

Acute-phase behavior of factor VIII procoagulant and other acute-phase reactants in rabbits

We measured the plasma concentrations of factor VIII procoagulant (FVIII:C) in rabbits in two laboratory models of inflammation and after injections of purified homologous interleukin 1 (IL-1). The mean FVIII:C activities 2-4 days following the intramuscular injection of turpentine were significantly elevated, as were the concentrations of fibrinogen and C-reactive protein (CRP). By contrast, rabbits given 50 ng of bacterial lipopolysaccharide intravenously developed increased levels of FVIII:C but not of fibrinogen or CRP. We then studied plasma levels of FVIII:C, fibrinogen, and CRP after injections of either form of purified rabbit IL-1. If the rabbits' temperatures were monitored, FVIII:C levels increased in a log dose-related manner. This effect was not seen unless the rabbits were restrained and was seen in restrained rabbits even if the temperatures were not monitored. Fibrinogen and CRP levels did not change after IL-1 injections. These findings demonstrate that FVIII:C is an acute-phase reactant as judged by its response to turpentine inflammation or endotoxin injections, and to injections of IL-1 in restrained rabbits. FVIII:C appears to be a more sensitive acute-phase reactant than either fibrinogen or CRP. Neither of the latter proteins responded to low-dose endotoxin injections nor to either form of purified homologous IL-1. The doses of IL-1 given did cause fever, neutrophil leukocytosis, and hypoferremia.

Increased Serum Antibacterial Activity After Terpentine-Induced Acute Inflammation

An acute inflammatory response was induced in rabbits by an intramuscular injection of turpentine. After this injection serial serum samples were obtained and serum haptoglobin levels were monitored. In addition, increased antibacterial activity was measured using a viable plate and photometric growth assay. As early as 4 hrs after turpentine challenge an increase was noted in serum antibacterial activity towards Staphylococcus aureus. At 48 hrs pronounced killing activity was demonstrated against S. aureus and S. epidermidis but not against Escherichia coli or other gram negative bacilli. By five days another serum bactericidal activity was present against Bacillus subtilis. Enhanced serum antibacterial activity persisted for 2 weeks. Partial characterization of these factors indicates that they are probably beta lysins.

Control of the acute phase response. Demonstration of C-reactive protein synthesis and secretion by hepatocytes during acute inflammation in the rabbit.

To determine the cell of origin of C-reactive protein (CRP) and to cast light on the mechanisms leading to the acute phase response, we used an immunoenzymatic technique to visualize this protein in livers from rabbits at intervals after intramuscular injection of turpentine. CRP was detected only in hepatocytes. 8 h after turpentine injection, CRP was demonstrated in occasional periportal hepatocytes. With time, larger numbers of positive cells were detected successively in perilobular, midlobular, and centrilobular areas. On electron microscopy, CRP was detected in rough endoplasmic reticulum (RER), smooth endoplasmic reticulum (SER), and Golgi apparatus (GA). When colchicine was administered to inhibit cellular secretion of CRP, intensity of reaction and number of CRP-containing hepatocytes were substantially greater than without colchicine, but the sequence of intralobular distribution was similar. At peak serum response 38 h after turpentine injection, CRP could be demonstrated in most hepatocytes. Electron microscopic studies showed accumulation of CRP on membranes and lumina of RER, SER, GA, and in cytoplasmic vacuoles. These findings indicate that CRP is produced by progressively increasing numbers of hepatocytes after inflammatory stimulus and suggest that a mediator, acting initially in portal zones, is responsible for recruitment of cells to CRP production.

Effect of inflammation on erythroid precursors (BFU-E and CFU-E) in bone marrow and spleen of mice.

Inflammation, induced in mice by a single intramuscular injection of turpentine, caused a long lasting reduction in the number of morphologically unrecognizable CFU-E in the bone marrow. This resulted in marked decreases in marrow erythroblasts and their iron incorporation into heme. The effect is attributed to blood-borne mediators of inflammation which either caused an emigration of progenitors of CFU-E from the marrow or inhibited their proliferation, possibly as the result of increases in marrow myelopoiesis. CFU-S as well as erythroid and myeloid precursors were markedly increased in the spleen during the inflammatory reaction.

Increased bilirubin formation from nonhemoglobin sources in rats with disorders of the liver.

Abstract The hepatic component of the early labeled fraction of bile pigment was found to be increased in rats under the following conditions: toxic liver damage after repeated injections of carbon tetrachloride, liver regeneration from 2 to 5 days after subtotal hepatectomy, treatment with hydrocortisone or phenobarbital, and infllammatory disease due to intramuscular injection of turpentine. The rise in labeled bilirubin production in these experiments was qualitatively different from that observed with accelerated erythropoiesis, consisting of enlargement of the initial sharp component rather than the plateau phase of the early pigment fraction. By contrast, formation of early labeled bilirubin was significantly depressed in 2 of 3 rats studied within a few hours after subtotal hepatectomy. These experiments suggest that increased bilirubin formation from nonhemoglobin sources may occur under a variety of pathologic and pharmacologic conditions and is a potential mechanism of hyperbilirubinemia in liver disease.

CLINICAL AND EXPERIMENTAL EFFECT OF COBALT ON VARIOUS TYPES OF ANEMIA (PART 1)

As the basis of treatment of anemia by cobalt, experimental anemia in rabbits was used, and compared with the effect of cobalt and that of iron.Cobalt produced excellent effect than iron on the infectious anemia, associated with sterile inflammation induced by the intramuscular injection of turpentine in the rabbits. Also cobalt produced similar effect on the anemia, due to staphylococcal infection by the intramuscular injection. But there is no effect on the phenylhydrazine anemia and less effect on anemia due to bleeding.The daily oral dosage of 30-60 mg of cobaltous chloride administered to 35 patients with various types of anemia produced erythropoetic responses in 10 cases.In 6 of 10 patients with iron deficiency anemia, in 2 of 8 patients with infectious anemia, and 1 patient with inoperable sarcoma of stomach, definite reticulocytic responses and increase in red cells, hemoglobin and hematocrit values were observed.But in cases with refractory anemia, chronic nephritis, thrombocytopenic purpura, and leukemia no effect were observed.Effect of cobalt on the iron refractory anemias, especially on infectious anemia and anemia due to tumor was discussed.