Analysis of flumethasone pivalate formulations by high-performance liquid chromatography.
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Biomedical subjects
Publications and source records attributed to A Vincent.
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We have investigated the organization and transcription of several genes in Drosophila melanogaster which are clustered on an 18-kb cloned DNA fragment (c25) that maps at 99D on the cytogenetic map. Multiple mRNAs, transcribed from genes which lie adjacent to a ribosomal protein ( rp49 ) gene, are present during oogenesis, embryogenesis, or both. At least five mRNAs are transcribed from one of these genes ( EH8 ); three are zygotic transcripts, of which two are blastoderm stage-specific, whereas two others accumulate during oogenesis and are therefore matenal mRNAs. The complex transcription pattern of this gene indicates that alternate usage of protein-coding exons results in the production of different mRNAs with different coding capabilities during oogenesis and embryogenesis. The EH8 transcription unit is framed by genes actively expressed in the adult male fly; thus, the blastoderm stage-specific promoter may be silent although within a region of transcriptionally active chromatin.
The nonspecificity and variety of clinical features in acute pulmonary embolism account for a high suspicion rate, with a further exclusion of the diagnosis in up to 80% of cases. Arterial blood gas analysis and other laboratory tests are highly non-specific, and suggestive signs on the electrocardiogram are uncommon. The plain chest X-ray often yields valuable information. Lung scan, when normal, has the great advantage of excluding pulmonary embolism. At present, no procedure has superseded pulmonary angiography to attest the presence of emboli. Angiography will probably gain widespread acceptance with the larger use of thrombolysis.
Mice were injected for 1-2 months daily with 10 mg immunoglobulin G (IgG) from four patients with Lambert-Eaton myasthenic syndrome (LEMS); control mice were injected with pooled human IgG from normal donors. Gastrocnemius muscles were homogenised for the assay of acetylcholine (ACh), choline acetyltransferase (ChAT), and cholinesterase (ChE). The ACh, ChAT, and ChE contents of gastrocnemius muscles from "LEMS mice" were about the same as the control values, which were 180 pmol, 40 nmol X h-1 (37 degrees C), and 15 mumol X h-1 (37 degrees C), respectively. Hemidiaphragms were treated with an irreversible ChE inhibitor (Soman) and incubated at 20 degrees C for estimation of ACh release. Resting ACh release from experimental muscles was reduced by about 25% (P2 less than 0.05) and the release evoked by 3 s-1 nervous stimulation by 50% (P2 less than 0.05). On the other hand, 50 mM KCl-induced transmitter release was not abnormal in LEMS mice. The findings indicate that IgG antibody from patients with LEMS may bind to nerve terminal determinants that are involved in quantal and nonquantal ACh release.
Two stored frozen massive bone allografts were implanted after resection of femoral and tibial tumours in two patients. Eighteen and thirty months later the authors had the opportunity to make a microscopic examination of the grafts. The cortical bone grafts were incorporated by the classic "creeping substitution." The repair of the cortical transplant was very incomplete and showed large resorption cavities. The newly-formed living bone was not fully mineralized. These phenomena related only to the outer area of the cortical bone, the dead intra-cortical area being left unaffected by the process. Cancellous bone graft repair was faster and more efficient.
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In vivo protein synthesis in duck erythroblasts was compared to in vitro translation of polyribosomal and free cytoplasmic mRNA. The in vivo study showed the absence of de novo synthesis of the Mr 73 000 poly(A)-binding protein found associated with all polyribosomal mRNA. In vitro translation demonstrated that the mRNA for this protein is absent from the polyribosomal mRNA fraction but constitutes a medium frequency messenger among the repressed free mRNA. This result confirms the existence of a qualitative translational control in terminal differentiating duck erythroblasts leading eventually to the arrest of the protein synthesizing machinery.
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Miniature end-plate potential (mepp) amplitudes were investigated in mouse diaphragm exposed in vitro to different serum fractions from seven patients with myasthenia gravis who had elevated serum anti-acetylcholine receptor antibody levels and from controls. The mepp amplitudes were significantly reduced by whole myasthenic sera, restored by washing, and not reduced by heated (56 degrees C) myasthenic sera, which would inactivate complement but not antireceptor antibody. Immunoglobulin G (IgG)-depleted myasthenic sera also significantly reduced mepp amplitudes, while the IgG fraction alone or with normal serum did not. The results indicate that in vitro reduction of mepp amplitudes in mouse muscle by myasthenic sera is not dependent on the IgG fraction alone, and requires a heat-sensitive factor.
Two types of in vivo untranslated 'free' mRNA-protein particles (mRNP) were isolated from duck erythroblast cytoplasm and characterised. Both types, namely the highly purified globin mRNA-specific '20S' mRNP and the '35S' mRNP containing a heterogenous non-globin mRNA population, are not translatable in rabbit reticulocyte lysates, but yield active mRNA upon deproteinisation. In vivo, 90% of globin mRNA is translated, but the majority of mRNA types are found in the inactive mRNP fraction, including fully repressed mRNA species. Searching for the factors controlling differential mRNA repression, we characterised and compared the protein composition of globin and '35S' mRNP using two dimensional gel electrophoresis, in vivo labelling with [35S]methionine and in vivo phosphorylation. The major proteins ubiquitously bound to globin or any other mRNA in the polyribosomes (e.g., the 73 K mol. wt. poly(A) binding protein) were not detected in purified inactive mRNP. In the latter some polypeptides appear to be associated with only one of the two inactive mRNA types while some others are common to both mRNPs. Furthermore, different rates of synthesis and phosphorylation characterize the protein populations of the two types of repressed mRNP. The specificity in composition and metabolism of the populations of polypeptides associated with different subpopulations of inactive cytoplasmic mRNA, as shown here, argues in favour of a role of mRNP proteins in mRNA recognition and selective translational repression, possibly in association with the ScRNA previously found as components of the free mRNP and able to inhibit protein synthesis.
Anti-acetylcholine receptor (anti-AChR) antibodies from two myasthenia gravis (MG) patients, known to have anti-AChR to the alpha-bungarotoxin (alpha-BuTx) binding site on the AChR (anti-alpha-BuTx site), were fractionated on an affinity column of human AChR fully saturated with alpha-BuTx. More than 80% of the anti-AChR in the pass-through fractions was directed against the alpha-BuTx site, whereas anti-AChR eluted from the column was mostly directed at other sites on the AChR. Recovery of anti-AChR was greater than 60%. Anti-alpha-BuTx site antibodies varied from 0 to 33% of the total anti-AChR antibodies in 12 MG sera. Fractionation on Sepharose-Protein A showed that anti-alpha-BuTx was only restricted to IgG3 in two patients. Anti-alpha-BuTx site antibody, which can be separated from antibodies binding to other antigenic determinants on the AChR, is variable in amount and heterogeneous in its subclass distribution. Although it may play an important pathogenic role in some patients, our results do not support the existence of an anti-alpha-BuTx site antibody common to all MG patients.
Recent years have seen considerable progress in understanding the nature of the molecular events involved in neuromuscular transmission. The acetylcholine receptor (AChR) has been purified to homogeneity and acetylcholine-induced ion transport has been reconstituted by incorporation of pure AChR into artificial membranes. Immunization against purified AChR induces a condition, clinically and physiologically similar to the human disease myasthenia gravis, which is due to circulating anti-AChR antibodies. This model, experimental autoimmune myasthenia gravis, is proving useful for investigating the role of genetic factors in determining the immune response to AChRs and for testing various experimental approaches to specific treatment. Myasthenia gravis is an autoimmune disease in which there is loss of acetylcholine receptors at the neuromuscular junction. Anti-AChR antibodies can be detected in the majority of patients and they cause loss of AChR by a variety of mechanisms. Anti-AChR antibody is heterogeneous and not restricted in idiotype. The role of the thymus in MG is still uncertain, but recent experiments implicate the presence of a cell type in MG thymus which may be involved in autosensitization to AChR.
In the Lambert-Eaton myasthenic syndrome (LEMS), there is a decreased release of acetylcholine quanta from the nerve terminal by nerve impulse. Recently, an autoimmune origin of LEMS was documented by passive transfer of its electrophysiologic features from man to mouse with IgG. Freeze-fracture electron microscopy of LEMS neuromuscular junctions has revealed a paucity of presynaptic membrane active zones. Thus, the active zones might be the targets of the pathogenic autoantibodies in LEMS. To test this assumption, freeze-fracture electron microscopic studies were done in mice injected with 10 mg of IgG daily from each of three LEMS patients and in control mice treated with normal human IgG or no IgG. IgG from patients 1 and 2 impaired neuromuscular transmission in mice, but IgG from patient 3 failed to do so. After 52-69 days of treatment, diaphragm or anterior tibial muscles were removed and coded. Paired muscles from control mice and mice receiving LEMS IgG were studied "blindly." Satisfactory freeze-fracture replicas of 185 presynaptic membrane P-faces were analyzed by stereometric methods. In mice treated with LEMS IgG that was pathogenic by electrophysiologic criteria, there was a selective depletion of active zones and active-zone particles but not of other membrane particles and there was a concomitant increase of large membrane particles aggregated into clusters. These findings provide additional evidence that the active zones facilitate quantal transmitter release by nerve impulse, lend further support to the assumption that the active-zone particles are Ca2+ channels, and establish mediation of the membrane lesions in LEMS by IgG.
The in vivo degradation of endplate acetylcholine receptors was investigated in mice treated daily with IgG from 10 myasthenia gravis patients. Four preparations increased the rate of degradation. The number of endplate acetylcholine receptors after 7 days of IgG treatment was greater than that predicted on the basis of the increased degradation rate, suggesting a compensatory increase in receptor synthesis.
The in vivo synthesis of diaphragm endplate acetylcholine receptors was estimated in mice treated daily with IgG from eight myasthenia gravis patients. Myasthenia gravis IgG preparations which had previously been shown to increase the rate of receptor degradation also increased the rate of receptor synthesis, suggesting the existence of a compensatory mechanism serving to stabilise the number of acetylcholine receptors.
We studied serum anti-acetylcholine receptor (AChR) antibody and clinical response to thymectomy in myasthenia gravis for 1 to 3 1/2 years postoperatively in 25 patients who did not receive immunosuppressive drugs. Clinical grade was assessed "blind." Mean final anti-AChR values were significantly reduced compared with thymectomy values (69.6 +/- 7.5% SEM; p less than 0.05). Anti-AChR fell steadily to 42-15% in the six patients who developed remission. Overall, there was a significant correlation between changes in anti-AChR and in clinical grade at 1 year (p less than 0.01) and at final assessment (p less than 0.001). An association between fall in anti-AChR and clinical improvement was absent in five individuals and not accounted for by change in antibody characteristics.
Thirty patients with a painful hip arthroplasty had an In-111 leucocyte scan before surgical reexploration. In 12 patients, the In-111 leucocyte scan was abnormal and in all of them, microorganisms were found at the culture of the material from their hips at the operation. Among the 18 patients with a normal scan no infection was found in 17. In one patient, a thick-walled abscess growing Escherichia coli was found. We conclude that In-111 scanning is sensitive, specific and therefore useful in the differential diagnosis of pain after hip arthroplasty.