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[Results of intramuscular injection of glycosamino-glycanpolysulfate (GAGPS) in experimental arthrosis of the knee in dogs (author's transl)].

Report on the action of GAGPS injected intramuscularly on experimental arthrosis of the knee-joint in dogs. In both hind-legs the lateral tibial joint-surface and the lateral meniscus were resected. This produces arthrotic changes similar to human arthrosis within 2 months. Of 27 dogs 13 received i.m. injections into the glutei of GAGPS (25 mg/kg). 14 dogs were not untreated controls. After the 7th postoperative day the injections were given on every third day on 10 occasions, thereafter every fourth day for another 10 occasions. The animals were killed 3 months after the start of the experiment. Loss of cartilage on the lateral condyle proved to be clearly less pronounced in the treated animals than in the controls, macroscopically, radiologically and histolocally. GAGPS, injected, intragluteally, limits loss of cartilage in experimental arthrosis of the knee in dogs.

Animals↗

Intramuscular injection of plasmid DNA expressing mRNA7 coding the nucleocapsid protein of JHMV partially protected mice against acute infection with JHMV.

We constructed a plasmid expressing mRNA7 coding the nucleocapsid (N) protein of JHM strain of mouse hepatitis virus (JHMV) under the control of Rous sarcoma virus LTR, referred to as pRSV-mRNA7. When C57BL/6 mice injected intramuscularly (i.m.) with control plasmid DNA which contained no viral sequence were infected with JHMV, necrotic figures of neural cells and diffuse immersion of lymphatic cells in the pedunculus cerebri were observed. In the hypothalamus, vascular cuffing consisting of lymphatic cells was observed. In contrast, no histological change was observed throughout these areas of the brains in the JHMV-infected mice after injection with pRSV-mRNA7. These results showed that the injection with plasmid DNA expressing mRNA7 of JHMV partially protected mice against acute infection with JHMV in the brain. The plasmid DNA was i.m. injected into mice and the cytolytic activity of spleen cells from the mice was assessed by 51Cr-release assay. The spleen cells from the mice administrated with pRSV-mRNA7 showed a significant level of cytolytic activities against the transfected cells expressing the viral N protein even though the spleen cells were not cocultivated with stimulator cells. When the spleen cells from administrated mice with pRSV-mRNA7 were cocultivated with stimulator cells, higher cytolytic activity was observed against the transfected cells, compared with the activity without stimulation.

Acute Disease↗

[Therapeutic angiogenesis through intramuscular injection of the gene for vascular endothelial growth factor (VEGF)].

Therapeutic angiogenesis by injection of growth factors or genes encoding for these appears a promising strategy for treatment of critical limb ischaemia but is currently still in an experimental phase. Among the cytokines that promote angiogenesis, i.e. the postnatal growth of blood vessels due to activation and proliferation of endothelial cells, vascular endothelial growth factor (VEGF) takes a special position. VEGF has proven to be a powerful angiogenic factor in vivo that specifically promotes migration and replication of endothelial cells. The specificity of VEGF for endothelial cells, compared with the more widespread effects of other factors such as fibroblast growth factor (FGF), may provide a broader therapeutic potential. The recombinant form of the human VEGF gene can be injected intramuscularly as a plasmid (phVEGF). The angiogenic effect of phVEGF can be explained in several ways: by direct stimulation of proliferation and migration of endothelial cells in vivo; by recruitment of previously existing collaterals; and by improving vasomotor function of vessels in ischaemic tissue.

Animals↗

Treatment of cystic ovarian disease in dairy cattle. Comparative observation of the effects of an intramuscular injection of corticosteroids and an intravenous injection of a combination of human chorionic gonadotropin and progesterone.

Of 67 cows with cystic ovarian disease (COD), 34 were injected intramuscularly with 20 mg of betamethasone of 10 mg of dexamethasone (CC) and 33 intravenously with a combination of 3,000 IU of human chorionic gonadotropin and 125 mg of progesterone (HCG -P). Pregnancy rates and intervals between treatment and conception were 32.4% and 32.1 +/- 30.5 days, respectively, in the CC-treated cows and 30.3% and 51.6 +/- 29.5 days in the HCG -P-treated ones. Cows which had not responded to gonadotropin treatment showed a considerably higher pregnancy rate when treated with CC and a lower pregnancy rate when administered with HCG -P than those which had received no treatment before. There was a trend that the earlier a cow was treated, the more readily she recovered. CC injection gave a satisfactory result even when performed long after calving. The estrous behaviour seemed to be related with prognosis in cows with COD. In both CC-treated and HCG -P-treated cows, the pregnancy rate was the highest in cows showing anestrus and the lowest in those exhibiting irregular estrus.

Animals↗

Induction of apoptosis in lymphoid tissues of mice after intramuscular injection of enterotoxigenic Escherichia coli enterotoxin.

Although it has been reported that intravenous injection of Escherichia coli enterotoxin induces atrophies of the thymus and spleen by necrosis, the toxin injected intramuscularly to mice induced atrophies of both tissues, which were associated with apoptosis of lymphocytes. Apoptosis predominantly occurred in the thymus and increased in a time-dependent manner up to 26 h and faint ladder band patterns of DNA were observed at 36 h. Although the high dose of toxin also induced in vitro apoptosis in cultured thymocytes, the toxin was not detected in the serum at levels sufficient to cause in vitro apoptosis after intramuscular administration. By flow cytometric analysis, CD4+ CD8+ double-positive T cell and CD45+ positive B cell numbers were found to be mainly decreased in thymus and spleen, respectively, of mice. These results suggest that the atrophies of thymus and spleen by intramuscular administration of the toxin to mice are due to apoptosis of CD4+ CD8+ double-positive T and CD45+ positive B cells, respectively, but the toxin does not reach these cells via the circulation. A different mechanism from that in vitro in cultured cells might be involved in the induction of apoptosis in vivo.

Adjuvants, Immunologic↗

Serum pharmacokinetics of oxytetracycline in sheep and calves and tissue residues in sheep following a single intramuscular injection of a long-acting preparation.

The pharmacokinetics of a long-acting oxytetracycline (OTC) formulation (Liquamycin LA-200) injected intramuscularly (i.m.) at a dose of 20 mg/kg were determined in four calves and 24 sheep to determine if the approved label dose for cattle provided a similar serum time/concentration profile in sheep. The AUC for the calves was 168+/-14.6 (microg ? h/mL) and was significantly less than the AUC for sheep (209+/-43 microg ? h/mL). Using the standard two-stage approach and a one-compartment model, the mean Cmax for the calves was 5.2+/-0.8 microg /mL, and for the sheep was 6.1+/-1.3 microg /mL. The mean terminal phase rate constants were 0.031 and 0.033 h, and the Vdss were 3.3 and 3.08 L/kg for the calves and sheep respectively. Analysis of the data using the standard two-stage approach, the naive pooled-data approach and a population model gave very similar results for both the cattle and sheep data. Sheep tissue residues of OTC in serum, liver, kidney, fat, muscle and injection site were measured at 1, 2, 3, 5, 7 and 14 days after a single i.m. injection of 20 mg/kg OTC. Half-lives of OTC residues in the tissues were 38.6, 33.4, 28.6, 25.4, 21.3, and 19.9 h for injection site, kidney, muscle, liver, mesenteric fat and renal fat, respectively. The ratio of tissue to serum concentration was fairly consistent at all slaughter times, except for the fat and injection sites. The mean ratios were 1.72, 4.19, 0.11, 0.061, 0.84 and 827 for the liver, kidney, renal fat, mesenteric fat, muscle and injection sites, respectively. The tissue concentrations of OTC residues were below the established cattle tolerances for OTC in liver (6 p.p.m.), muscle (2 p.p.m.) and kidney (12 p.p.m.) by 48 h, and in injection site muscle by 14 days after the single i.m. injection of 20 mg/kg.

Animals↗

Steroid levels after intramuscular injection of testosterone propionate in the caprine.

Nine sexually mature intact grade does were injected intramuscularly with testosterone propionate and subsequent plasma steroid concentrations determined and male-like behavior recorded. The does received either 100 mg testosterone propionate every three days for six treatments, total dose 600 mg (N = 5); 50 mg testosterone propionate daily for eighteen days, total dose 900 mg (N = 2) or 10 mg testosterone propionate daily for eighteen days, total dose 180 mg (N = 2). The treatments induced male-like sex behavior, the intensity of which was related to the dose of exogenous testosterone used, the regimen of administration, and the plasma levels of testosterone. Exogenous testosterone treatment had minimal effect on the subsequent reproductive activity of the does.

Animals↗

The effect of intramuscular injections of boar pheromone 5alpha-androstenol on the hormonal regulation of the estrous cycle in hypoosmatic gilts.

Until 1999 it was accepted that pheromones act exclusively by stimulating the dendritic receptors present in olfactory epithelium. Cycling gilts with an experimentally-disrupted neural olfactory pathway were used to test the hypothesis that boar pheromone 5alpha-androstenol may affect the secretion of hormones involved in the regulation of the estrous cycle by the humoral pathway. On day 12 of the estrous cycle the nasal cavity of gilts (n=15) was irrigated with zink sulfate solution. From day 16 to 20, the experimental group (n=10) was injected intramuscularly with 5alpha-androstenol (20 microg) twice a day. Blood samples were collected from the jugular vein at 4 h intervals on days 17-21 to estimate plasma concentration of LH, oxytocin, estradiol-17beta, testosterone and progesterone. The experimental group displayed a significantly lower mean concentration of LH than the control animals (P<0.0001). The decrease in concentration of LH was accompanied by the reduction of oxytocin (P<0.001), estradiol-17beta (P<0.001) and testosterone (P<0.01) secretion. These results demonstrated that 5alpha-androstenol influenced hormonal regulation by humoral pathway and might be considered to be the priming pheromone in gilts.

Androstenols↗

Remote intramuscular injection of immobilising drugs into fish using a laser-aimed underwater dart gun.

Sixty coldwater and warmwater fish ranging in weight from 2 to 35 kg were injected intramuscularly with the hypnotics alphaxalone-alphadolone and metomidate hydrochloride and the non-depolarising muscle relaxant gallamine triethiodide using a laser-aimed underwater dart gun. Alphaxalone-alphadolone produced sufficient sedation for easy netting within five to 20 minutes at doses between 0.3 and 0.5 ml/kg, with induction being somewhat faster in warmwater species. The pattern of induction was similar with metomidate but required doses of 40 to 60 mg/kg. The muscle relaxant gallamine triethiodide showed promise as a practical agent for the capture and handling of large fish by virtue of its smooth induction of paralysis at doses between 1 and 3 mg/kg and its reversible supplementation with orally administered metomidate hydrochloride.

Anesthetics↗

Plasma half-lives and bioavailability of human monoclonal Rh D antibodies BRAD-3 and BRAD-5 following intramuscular injection into Rh D-negative volunteers.

Two human MoAbs, BRAD-3 (an IgG3 anti-D) and BRAD-5 (an IgG1 anti-D), were produced from Epstein-Barr virus (EBV)-transformed B lymphoblastoid cell lines grown in hollow fibre bioreactors. Six Rh D-negative male volunteers were injected intramuscularly with anti-D; two received BRAD-3 (approx. 1500 micrograms, 2600 IU), two were given BRAD-5 (300 micrograms, 2000 IU), and two had polyclonal anti-D immunoglobulin (500 IU, approx. 100 micrograms anti-D). Levels of anti-D in plasma samples taken up to 42 days later were measured by a sensitive AutoAnalyser method. The half life of BRAD-5 (mean 22.2 days) was greater, and that of BRAD-3 (mean 10.2 days) less than that of polyclonal anti-D (mean 15.6 days). The bioavailability (plasma uptake) of the MoAbs (mean 33.9%) was less than that of the polyclonal anti-D (mean 60.3%). BRAD-3 and BRAD-5 may be suitable for use in antenatal and post-natal prophylaxis against Rh D haemolytic disease of the newborn.

Antibodies, Monoclonal↗

Identifications of radioactive steroid estrogen conjugates in blood plasma of laying hens after intramuscular injection of (4--14C)-estrone.

[4--14C] Estrone was injected intramuscularly into six laying hens. Fifty minutes later the hens were exsanguinated. The plasmas were examined for conjugates of radioactive phenolic steroids by recovery on columns of Amberlite XAD-2 or by extraction with tetrahydrofuran followed by chromatography on a column of DEAE-Sephadex A-25 in a gradient of NaCl. The biggest Sephadex chromatographic fraction (50,4% of total) contained about 42% of its radioactivity as estradiol-17alpha-3-sulfate and 18% as estradiol-17beta-3-sulfate and the remaining 40% was identified tentatively as estradiol-17alpha-17-sulfate plus a small proportion of estradiol-17beta-17-sulfate. The second biggest Sephadex chromatographic fraction (12.7% of total) was a mixture of conjugates not further identified. Minor fractions identified comprised estrone-beta-glucuronide (2.8%), estradiol-17alpha-3-beta-glucuronide (2.8%), estradiol-17beta-3-beta-glucuronide (2.3%) and estrone sulfate (6.0%). Evidence was obtained for the presence of small proportions of estradiol-17alpha disulfate and estradiol-17beta disulfate.

Animals↗

Steroid estrogen conjugates in hens' urine II. Identifications of some minor conversion products of intramuscularly injected [4(-14C)]estrone.

[4(-14C)]Estrone was injected intramuscularly into two mature laying Rhode Island Red hens. Radioactive steroids and steroid conjugates recovered from the urine on Amberlite XAD-2 columns were fractionated on columns (100 cm) of DEAE-Sephadex A-25 by NaC1 gradients. The presence of the following were confirmed, the figures in brackets indicating average proportions as per cent of total radioactivity recovered after Sephadex column chromatography:-the 3-beta-glucuronides of estrone (10.9) and of estradiol-17alpha plus estradiol-17beta(9.8): the 17-beta-glucuronides of estradiol-17alpha plus estradiol-17beta (2.1); the 3-sulfates of estrone (14.5) and of estradiol-17alpha plus estradiol-17beta(27.4); and the disulfates of estradiol-17alpha plus estradiol-17beta (2.3). The following additional conjugates were identified:-a beta-glucuronide of 16-epiestriol (0.2) and a beta-glucuronide of 16-keto-estradiol-17beta (0.2); the 3-sulfates of 16-epiestriol (1.4), of 17-epiestriol (0.9), of 16,17-epiestriol (0.7), of 16-keto-estradiol-17beta (1.1), and of 2-methoxyestrone (0.7). Some evidence was obtained for the presence of 16,17-epoxy-estratrienol-3-sulfate (1.9).

Animals↗

Direct intramuscular injection of plasmid DNA encoding angiopoietin-1 but not angiopoietin-2 augments revascularization in the rabbit ischemic hindlimb.

BACKGROUND: Angiopoietin-1 (Ang1) and angiopoietin-2 (Ang2) have recently been identified as ligands for the endothelial cell-specific Tie2 receptor. Little is known regarding the impact of these Tie2 ligands on postnatal neovascularization. Accordingly, we tested the hypothesis that gene transfer of plasmid DNA encoding Ang1 and Ang2 could modulate collateral vessel development in a rabbit model of hindlimb ischemia. METHODS AND RESULTS: pAng1* (n=15), pJFE control (no Ang1* insert) (n=9), pAng2 (n=9), pcDNA3 control (no Ang2 insert) (n=10), or saline (n=5) was injected intramuscularly into the rabbit ischemic hindlimb. Collateral vessel development and limb perfusion were assessed before and 30 days after treatment. Calf blood pressure ratio (ischemic to normal hindlimb) was increased 30 days after Ang1* gene transfer versus controls (Ang1*, 0.90+/-0.02; pJFE, 0.76+/-0.05; saline, 0.77+/-0. 03; P<0.05). Angiographic score was higher (P<0.05) in the pAng1* group (0.63+/-0.02) than in the pJFE (0.51+/-0.03) or saline (0. 52+/-0.02) group. Maximal (postpapaverine) blood flow in the ischemic limb was higher (P<0.05) after pAng1* (67.8+/-4.9 mL/min) than pJFE (51.2+/-4.4 mL/min) or saline (52.9+/-4.9 mL/min). Capillary density and capillary/muscle fiber ratio (242+/-12/mm2 and 0.89+/-0.06, respectively) were higher (P<0.01) with pAng1* than pJFE (172+/-11/mm2 and 0.64+/-0.05) or saline (166+/-10/mm2 and 0. 67+/-0.05). Neovascularization was not enhanced with pAng2. CONCLUSIONS: Ang1 but not Ang2 gene transfer produces anatomic and physiological evidence of enhanced collateral vessel formation. Ang1 may modulate neovascularization in adult animals and thus represents a feasible therapeutic strategy for patients with tissue ischemia. The role of Ang2 in postnatal neovascularization remains to be clarified.

Angiography↗

Electric pulses increase the immunogenicity of an influenza DNA vaccine injected intramuscularly in the mouse.

Vaccination by intramuscular injection of naked DNA is very efficient in the mouse, but immunogenicity of DNA vaccines needs to be improved in man. The aim of our study was to determine in BALB/c mice if suitable electric pulses delivered to the muscle after DNA injection--a procedure called electrotransfer--could improve the immunogenicity of suboptimal doses of a DNA vaccine expressing the influenza hemagglutinin protein. The results show a significant enhancement of the cellular and antibody responses following electrotransfer for the 1- and 10-microg DNA doses, respectively, but no effect on a lower dose. At the 10-microg dose, the IgG and hemagglutination inhibition mean titres were increased 25-fold and the inter-individual variability was markedly reduced.

Animals↗

Cytolytic activity induced by intramuscular injection of plasmid DNA expressing the nucleocapsid protein of the JHM strain of mouse hepatitis virus into C57BL/6 mice.

We constructed plasmids expressing mRNA7 coding the nucleocapsid (N) protein of JHM strain of MHV (JHMV) under the control of Rous sarcoma virus (RSV) LTR or human elongation factor (EF) 1a promoter, referred to as pRSV-mRNA7 and pEF-mRNA7, respectively. Although only a slight level of cytolysis was observed by the spleen cells from C57BL/6 mice injected intramuscularly with pEF-mRNA7, the spleen cells from the mice administered with pRSV-mRNA7 showed a significant level of cytolytic activities against the cells expressing the viral N protein. The difference in the level of specific cytolysis might have been mainly due to a difference in the expression levels of the N protein in the muscles between the mice injected with pEF-mRNA7 and pRSV-mRNA7, since the specific activity of chloramphenicol acetyltransferase (CAT) in muscles from the mice injected with plasmid DNA expressing CAT gene directed by RSV LTR was significantly higher than that in those administered by the plasmid DNA directed by EF-1a promoter.

Animals↗

[Specific cellular and humoral immune responses induced by intramuscular injection of DNA vaccine containing HBV HBsAg gene in mice].

To know the influences on the function of cellular and humoral immunity in mice by DNA vaccine containing HBV HBsAg gene, the DNA vaccine, pcDNA3-HBsAg, in which the HBsAg gene sequences of HBV were inserted into downstream of CMV promoter in pcDNA3, were injected intramuscularly into Balb/C mice. The results were that the serum anti-HBsIgG was detected in all of the pcDNA3-HBsAg immunized mice, but none in control mice; serum levels of Th1 cytokines, interleukine 2, and interferon gamma, were significantly higher in pcDNA3-HBsAg immunized mice than these in controls. It is indicated that the good responses of cellular and humoral immunity can be induced by injection of pcDNA3-HBsAg in Balb/C mice.

Animals↗

Treatment of ovarian cyst in swine by intramuscular injection with luteinizing hormone-releasing hormone analogue.

The diagnosis of ovarian cyst was made in 53 anestrous sows, which were treated experimentally with luteinizing hormone-releasing hormone analogue (LH-RH-A). Each sow was injected intramuscularly with 200, 400, or 500 micrograms one to five times. She was examined by rectal palpation in the course of treatment. The blood levels of estrogen and progesterone were estimated in three of the treated sows. From changes in them was followed the process of healing of ovarian cyst. As a result, (1) the interval between the beginning of treatment and the appearance of estrus was 26.9 days on the average, the rate of appearance of estrus 60.4%, that of healing of ovarian cyst 71.6%, that of conception 70.2%, and the number of young of a littler 9.0 on the average. (2) In the three sows subjected to estimation, the blood progesterone concentration increased after injection with LH-RH-A to reach almost the same level as that estimated in the corpus-luteum stage. The time of increase after injection was almost identical with the time when the luteinization of ovarian cyst was found by palpation. In a sow which manifested normal estrus, no ovulation took place, but a cyst was formed in the ovary and the blood estrogen concentration maintained at a high level for 10 days after the beginning of estrus.

Animals↗