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At least 19 recordsLinked to original sources

Myosin regulatory elements as vectors for gene transfer by intramuscular injection.

Intramuscular injection of plasmid constructs promises to be an effective way of carrying out gene therapy for muscle disorders as well as using muscle as an in vivo expression system for disorders that involve the gene product being secreted into the bloodstream. The effectiveness of this method depends on the design of the cassette used for the expression of the cDNA of the introduced gene. We tested the levels of expression achieved by a number of muscle-specific promoters and a myosin light chain enhancer when spliced to the reporter gene chloramphenicol acetyltransferase (CAT), in vitro and in vivo by injection into fast and slow muscles of the mouse. The results show that the highest levels of expression are achieved by a combination of a truncated myosin heavy chain promoter and the enhancer, and that a whole range of expression levels is obtained with the other combinations tested. The data show that a cassette based on these elements should provide efficient vectors for the introduction and expression of genes following intramuscular injection of naked DNA.

Animals↗

Immune responses and protection obtained with rotavirus VP6 DNA vaccines given by intramuscular injection.

Intramuscular (i.m.) injection of murine VP6 DNA vaccines raised high titers of rotavirus-specific serum IgG and IgA antibodies in BALB/c mice. A Th1-like antibody response was generated based on the ratio of serum IgG2a to IgG1 antibodies. Rotavirus-specific serum IgA but not fecal IgA was detected in mice prior to rotavirus challenge. Partial protection against rotavirus challenge was achieved as measured by reduction of rotavirus antigen shedding in feces. A similar level of protection was found with a bovine rotavirus VP6 DNA vaccine against a murine rotavirus challenge, suggesting that heterologous protection can be obtained by immunizing with VP6 DNA vaccines. We did not directly test for cytotoxic T lymphocyte (CTL) activity, but in vivo depletion of CD8+ T cells in mice immunized with a murine VP6 DNA vaccine did not significantly change the duration of virus shedding or the pattern of protection obtained. This finding suggested that CD8+ CTL activity was not essential for the partial protection we obtained by i.m. immunization of mice with VP6 DNA vaccines.

Animals↗

[Intramuscular injections--an outdated form of administration? 6 cases of Staphylococcus aureus sepsis following intramuscular injections].

Intramuscular injections can lead to local and systemic complications, such as abscess and sepsis. These are often caused by Staphylococcus aureus, occur in immunocompromised as well as in immunocompetent persons, and often need extensive medical and surgical treatment. We describe 6 cases with sepsis and multiple abscesses caused by Staphylococcus aureus after intramuscular injections. In view of possible serious complications, the indication for intramuscular injection as a method of drug administration is critically analyzed.

Abscess↗

Control of erythropoietin delivery by doxycycline in mice after intramuscular injection of adeno-associated vector.

We reported previously that controlled expression of a foreign gene in response to tetracycline derivative can be accomplished in mice by the autologous transplantation of retrovirus-modified muscle cells. Although regulated systemic delivery of therapeutic proteins from engineered tissues has potential clinical application, the transplantation of muscle cells is not currently feasible in humans. Several studies have shown that a single injection of adeno-associated virus (AAV) vectors into mouse muscle results in long-term expression of reporter genes as well as sustained delivery of proteins into the serum. Because this method is potentially applicable clinically, we constructed an AAV vector in which the expression of the mouse erythropoietin (Epo) cDNA is modulated in response to doxycycline. The vector was injected intramuscularly in normal mice. We observed that hematocrit and serum Epo concentrations could be modulated over a 29-week period in response to the presence or absence of doxycycline in the drinking water of these animals. Thus, a regulated gene expression cassette can be incorporated into a single AAV vector, such that intramuscular injection of the vector allows sustained and regulated expression of a desired gene.

Animals↗

Efficacy of 3,4,3-LI(1,2-HOPO) for decorporation of Pu, Am and U from rats injected intramuscularly with high-fired particles of MOX.

This study aimed to assess the efficacy of 3,4,3-LI(1,2-HOPO) for reducing uranium, plutonium and americium in rats after intramuscular injection of (U-Pu)O2 particles (MOX). Sixteen rats were contaminated by intramuscular injection of a 1 mg MOX suspension and then treated daily for 7 d with LIHOPO (30 or 200 micromol kg(-1)) or DTPA (30 micromol kg(-1)). LIHOPO was inefficient for removing Pu, Am and U from the wound site. However, it reduced Pu retention in carcass and liver by factors of 2 and 6 respectively, and Am retention in carcass and liver by factors of 10 and 30. In contrast, the effect of LIHOPO on U was to decrease the retention in kidneys by a factor of 75. These results confirm that LIHOPO is a good candidate for use after contamination with MOX, in combination with localised wound lavage or surgical treatment aimed at removing most of the contaminant at the wound site.

Americium↗

Lymphadenitis in experimental murine toxoplasmosis induced by intramuscular injection of tachyzoites.

When tachyzoites (RH strain) of Toxoplasma gondii are injected intramuscularly, experimental mice survive up to 7 days, 1-2 days longer than those infected intraperitoneally. We observed sequential histopathological changes in inguinal lymph nodes after intramuscular injection of tachyzoites to thighs of specific pathogen free (SPF) mice. Initial findings on 1 or 3 days after the injection were reactive germinal centers, distended sinuses and epithelioid cell clusters in cortical and paracortical regions. Later on 5 days after the injection, however, effacement of nodal structure with depletion of cells and focal necrosis were observed. Necrotizing lymphadenitis in the experimental murine toxoplasmosis suggests the causal relation between T. gondii infection and the human disease.

Animals↗

Tissue damage and concentration at the injection site after intramuscular injection of chemotherapeutics and vehicles in pigs.

Intramuscular injection sites were examined for macroscopical and microscopical changes and for residues of drugs six and 30 days after injection of chemotherapeutic preparations or vehicles in swine. The chemotherapeutic preparations contained sulphonamide and/or trimethoprim. All the chemotherapeutic preparations and the vehicles except physiological saline and sterile water caused macroscopical and microscopical changes, mainly appearing as areas of necrotic muscle tissue six days after the injection and as scar tissue 30 days after the injection. Residues of drugs were found at nearly all the injection sites six days after the injection, while 30 days after the injection only residues of sulphonamides were detectable in nearly half of the injection sites.

Animals↗

Selection of sites for intramuscular injections in the neck of the horse.

A radio-opaque marker was injected intramuscularly at different levels and with needles of different lengths into the cervical musculature of a series of ponies and horses which were under profound anaesthesia prior to euthanasia. Subsequently the necks were sectioned and radiographed to determine the fate of the injected material. The results indicated that care was necessary in the selection of the level for injection in order to prevent deposition of material on to the periosteal surface of the cervical vertebrae or on to the ligamentum nuchae and fascial sheaths. The muscle mass over the third vertebra was relatively poor but increased in the more caudal cervical regions. The best site for intramuscular injection appeared to be at the level of the fifth cervical vertebra, ventral to the funicular part of the ligamentum nuchae but dorsal to the brachiocephalic muscle. Even at this position injection material sometimes entered intermuscular fascial sheaths.

Animals↗

Diffusion and fate of intramuscularly injected human rabies immune globulin.

The importance of rabies immune globulin (RIG) in postexposure rabies treatment is well known and it has been emphasized that the local injection into the animal bite sites is crucial. This preliminary study used a radioisotope tracer that allows following the fate of human rabies immune globulin (HRIG) injected intramuscularly. There was significant retention and local diffusion of the immune globulin at the injection site and significant radiotracer could still be detected at the site 24 h later.

Animals↗

Copper and zinc deficiencies treatement by intramuscular injections in sheep.

In two experiments, compared to 10 control, 10 ewes were injected intramuscularly either with metallic copper dust or zinc oxide, suspended in purified olive oil. Copper injection raised the plasma and hepatic copper of the treated ewes. Similarly, injected zinc oxide produced a rise in plasma zinc. These increases started respectively on the 6th and 3rd day after the injection and lasted till the 21st or 17th day. No detrimental inflammation appeared. This form of treatment is therefore efficient against trace element deficiencies without any side effect.

Animals↗

Antigen-specific antibodies in cerebrospinal fluid after intramuscular injection of ovalbumin in horses.

Eighteen normal horses were assigned to 1 of 3 treatment groups to investigate the effects of IM or intrathecal (IT) administration of ovalbumin on serum and cerebrospinal fluid (CSF) antibody production. Horses of group 1 were injected intramuscularly with ovalbumin and adjuvant, while horses in treatment groups 2 and 3 received ovalbumin intrathecally or intravenously, followed by IM injection as in group 1. Serum and CSF antibody titers were tested in group I every 30 days for 4 months, while serum and CSF were collected in group 2 and 3 horses at postvaccination day 60. Horses of group 1 (IM) developed a serum antibody titer that peaked at postadministration day 60 (1:24,320 +/- 7,680) (mean +/- I SEM). Anti-ovalbumin antibodies were detected in CSF, and titers paralleled that of the serum, although at a much lower concentration (peak, 1:166 +/- 87). Horses of groups 2 and 3 developed significantly (P = .02) lower serum titers (1:720 and 1:2,067, respectively), but the difference in CSF titers did not achieve statistical significance (P = .06). The results confirm that antigen-specific antibody can be found in the CSF of horses in which antigen is not administered intrathecally. This may affect the interpretation of CSF analysis in diseases such as equine protozoal myeloencephalitis. Further, the findings suggest that IT injection of the soluble antigen ovalbumin induces a state of antigenic tolerance in the horse. The clinical significance of this finding remains unknown at this time.

Animals↗

Cell size and geometry of spinal cord motoneurons in the adult cat following the intramuscular injection of adriamycin: comparison with data from aged cats.

Adriamycin (ADM), an antineoplastic antibiotic, when injected intramuscularly, is taken up by motoneuron axonal terminals and retrogradely transported to the motoneuron soma where it exerts its neurotoxic effect. In the present study, ADM was injected into the hindlimb muscles of five adult cats. Measurements of the electrophysiological properties of the lumbar motoneurons innervating these muscles were obtained using intracellular techniques. Based upon these data the equivalent cylinder model of motoneurons was employed to evaluate ADM-induced changes in cell size and cell geometry. The size of cell somas in the ventral horn was also measured using light microscopy and computer imaging software. There were significant increases in the membrane time constant (25%) and input resistance (50%) in motoneurons whose muscles were treated with ADM (ADM-MNs) compared with data from control motoneurons (control-MNs). The increase in membrane time constant is attributed to an increase in membrane resistance; the increase in input resistance appears to depend upon both an increase in membrane resistance and a decrease in total cell surface area. Cell capacitance, which is proportional to the total cell surface area, was significantly reduced (15%) in ADM-MNs. Calculations based on cable theory indicate that while there was no significant change in the length of the equivalent cylinder for ADM-MNs, there was a significant decrease (17%) in the diameter of the equivalent cylinder. These data indicate that there is a decrease in total cell surface area which can be attributed to the shrinkage of branches throughout the dendritic tree. There was also a small (7%) but statistically significant decrease in the electrotonic length of ADM-MNs. Morphological analysis also revealed that the mean cross-sectional area of the somas of those ventral horn neurons which are likely to correspond to the motoneuron population was significantly reduced on the ADM-treated side compared to that of neurons on the control side. We conclude that significant geometrical changes were induced in lumbar motoneurons of adult cats after ADM was injected to their muscles. In old cats, spinal cord motoneurons exhibit similar patterns of changes in their electrophysiological characteristics which have also been suggested to be correlated with changes in cell geometry. The question then arises as to whether the response of motoneurons to ADM and the aging process reflects a stereotypic reaction of motoneurons to a variety of insults or whether the response to ADM mirrors specific aspects of the aging process.

Aging↗

Systemic correction of a fatty acid oxidation defect by intramuscular injection of a recombinant adeno-associated virus vector.

Mitochondrial beta-oxidation of fatty acids is required to meet physiologic energy requirements during illness and periods of fasting or physiologic stress, and is most active in liver and striated muscle. Acyl-CoA dehydrogenases of varying chain-length specificities represent the first step in the mitochondria for each round of beta-oxidation, each of which removes two-carbon units as acetyl-CoA for entry into the tricarboxylic acid cycle. We have used recombinant adeno-associated virus (rAAV) vectors expressing short-chain acyl-CoA dehydrogenase (SCAD) to correct the accumulation of fatty acyl-CoA intermediates in deficient cell lines. The rAAV-SCAD vector was then packaged into either rAAV serotype 1 or 2 capsids and injected intramuscularly into SCAD-deficient mice. A systemic effect was observed as judged by restoration of circulating butyryl- carnitine levels to normal. Total lipid content at the injection site was also decreased as demonstrated by noninvasive magnetic resonance spectroscopy (MRS). SCAD enzyme activity in the injected muscle was found at necropsy to be above the normal control mouse level. This study is the first to demonstrate the systemic correction of a fatty acid oxidation disorder with rAAV and the utility of MRS as a noninvasive method to monitor SCAD correction after in vivo gene therapy.

Animals↗

Concentrations of trimethoprim and sulphamethoxazole in the human prostate gland after intramuscular injection.

Following intramuscular injection of co-trimoxazole (trimethoprim/sulphamethoxazole 1 : 5), concentrations of trimethoprim (TMP) and sulphamethoxazole (SMX) were measured in the prostatic tissue and serum of 30 men undergoing endoscopic prostatectomy. Tissue levels for TMP appeared to be higher than serum levels and probably reached the minimum inhibitory concentration (MIC) for most urinary pathogens. Administration of the drug at 4, 8 and 12 h prior to sampling produced no significant difference in tissue levels. It is concluded that TMP is concentrated in prostatic tissue following intramuscular injection. Tissue levels for SMX were apparently lower than serum levels.

Anti-Infective Agents, Urinary↗

Comparison of liquid chromatographic and bioassay procedures for determining depletion of intramuscularly injected tylosin.

Crossbred pigs weighing 80-110 kg were injected intramuscularly in the ham with 8.8 mg/kg tylosin. Animals were slaughtered in groups of 3 at intervals of 4 h, and 1, 2, 4, and 8 days after injection, and samples of blood, injected muscle, uninjected muscle, liver, and kidney were analyzed by liquid chromatography (LC) and by bioassay using Sarcina lutea as the test organism. The LC method was far more sensitive with a detection limit of less than 0.1 ppm, while the detection limit by bioassay was about 0.5 ppm in tissue. Results by bioassay and LC sometimes differed considerably for tissue samples. Residues in all tissues were below the tolerance limit of 0.2 ppm at 24 h, except in the injected muscle in one animal. Residues were not detected in any tissue of any animal at 48 h after treatment.

Animals↗

Gene therapy of melanoma pulmonary metastasis by intramuscular injection of plasmid DNA encoding tissue inhibitor of metalloproteinases-1.

Tumor cell invasion and metastasis are a complex multistep process that involves the degradation of extracellular matrix proteins by matrix metalloproteinases. Tissue inhibitor of metalloproteinase-1 (TIMP-1) acts as a negative regulator of matrix metalloproteinases and thus prevents tumor cell invasion and metastasis by preserving extracellular matrix integrity. In the present study, we investigated whether increasing serum TIMP-1 levels by gene transfer would decrease experimental pulmonary metastasis of melanoma in C57BL/6 mice. Female animals bearing B16F10 melanoma pulmonary metastasis were injected intramuscularly twice per week with 100 microg of plasmid DNA encoding the human TIMP-1 cDNA (TIMP-1pDNA). Substantive levels of serum human TIMP-1 were observed 3 days after single injection and were found for 6 days thereafter. Pulmonary metastasis was significantly reduced in the mice following 4 weeks of TIMP-1 treatment as compared to the controls that were treated with the plasmid DNA vector alone. Further reduction of pulmonary metastasis and increase in survival were realized by intraperitoneal injection of 1000 U of IL-2 twice per week in combination with TIMP-1 treatment. In a parallel in vitro study, a 3-fold increase in TIMP-1 expression was observed in NIH3T3 cells after IL-2 treatment. Therefore, up-regulation of TIMP-1 expression by IL-2 likely contributed to the additive effect of IL-2 and TIMP-1 in reducing metastatic disease in the animal model. In conclusion, our findings support the potential of TIMP-1 gene therapy for the prevention of metastatic melanoma.

Animals↗