Can urethral pressure profilometry predict the response to colposuspension in bitches?
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Biomedical subjects
Publications and source records attributed to S P Gregory.
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We have compared the strength of a trout protamine gene promoter with that of the mouse beta major-globin gene by analysing the relative levels of run-off transcripts produced in a single mammalian in vitro transcription reaction. When the promoters are introduced on separate recombinant plasmids, the protamine transcripts are synthesised with much greater efficiency than those originating from the globin cap site. This enhanced transcription of the protamine gene is again observed when the promoters are applied as separate DNA fragments derived from the same recombinant plasmid. However, when the promoters are linked on a DNA fragment that includes 7 kb of DNA separating the initiation sites, then there is a marked reduction in the protamine signal relative to the globin. Deletion of a region of this fragment that contains the sequences flanking the globin gene at positions -335 to -1400 restores the enhanced protamine gene expression to the levels observed when the promoters are carried on separate DNA fragments.
The mRNA start site of a cloned rainbow trout protamine gene (TPG-3) has been localised using S1-nuclease mapping and primer extension of in vivo synthesised trout testis poly A+-RNA. The presumptive cap site occurs within an AT-rich region, only 14 nucleotides from the start of the protein-coding sequence. Transcription of this protamine gene in vitro, using the Hela whole-cell extract system, generates products initiated at the same nucleotide as that used in vivo. In vitro transcription is abolished by deletion of sequences between -20 and -48, within which is a canonical TATA-box having an llbp homology with the strong chick conalbumin and Adenovirus-2 major late promoters (CTATAAAAGGG).
A novel interaction between the 5'-untranslated region of eukaryotic messenger RNAs and non-contiguous sequences in the 18 S ribosomal RNA is proposed. The small ribosomal RNA contains, at its 3'-terminus, a heavily conserved hairpin structure. It is suggested that mRNA 5'-leader sequence stabilises this structure by interacting with other conserved nucleotides which flank it. Sequences closely related to the required sequence (A-U-C-C-A-C-C) occur quite commonly in eukaryotic mRNAs and are often found immediately upstream from the AUG-codon. This interaction may have a role in the events which lead up to the initiation of protein synthesis.
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Nuclei from Xenopus erythrocytes have been isolated under varying ionic conditions and their morphology studied by light and electron microscopy. The 200-A beaded chromatin fibre observed in vivo is maintained in media containing MgCl2 or CaCl2, but not in those containing Mn2+ as the sole divalent cation. In the absence of these alkaline-earth metal ions, 100-A filaments predominate. This suggests that the native 200-A fibre in isolated nuclei is also stabilized by specific divalent cations. Gross chromatin condensation within the nucleus occurs independently of whether the chromatin is in the form of 100-A or 200-A fibres, but requires the presence of divalent cations. Lower concentrations of MgCl2 or CaCl2 are necessary as compared with MnCl2. In the presence of sufficient divalent cations, decreasing KCl concentration causes peripheral condensation, the condensation of chromatin towards the nuclear membrane. Monovalent cation alone does not appear capable of inducing peripheral condensation. The transcriptional capacities of the different morphological types identified have been studied using E. coli RNA polymerase as a probe. Neither the degree of chromatin condensation nor the state of higher-order coiling have a significant effect on the rate of transcription of the DNA. These results are discussed with regard to further in vitro studies on eukaryotic gene activity.
The morphology of isolated Xenopus erythrocyte nuclei is largely determined by the ionic constitution of the isolation medium. Electron microscopy of thin sections of nuclei isolated under varying ionic conditions has revealed four different morphological types with characteristic chromatin configurations. These nuclei have been used to study the condensation of chromatin within the confines of the nucleus. Electron micrographs of thin nuclear sections have been analysed by computer on the basis of their intensity of staining with lead citrate/uranyl acetate. The staining pattern (image analysis scan) is characteristic for each nuclear morphology and can be explained by a model which assumes that there are only four higher-order conformations that chromatin can adopt within the nucleus. These are present in varying amounts in the different nuclear types and an attempt has been made to equate the peaks identified by image analysis with previously-described chromatin structures. The results of this study serve to emphasize the potential of computer-based image analysis in the study of chromatin structure.
Nuclei isolated from Xenopus erythrocytes can be transcriptionally reactivated by exposure to certain cytoplasmic proteins. The types of RNA synthesized during this reactivation have been studied and compared with those present in, or synthesized by, isolated nuclei not so reactivated or in entire Xenopus erythrocytes. In all cases, the pattern of transcription indicates the synthesis of a broad range of low molecular weight RNAs. Competitive hybridization demonstrates that the reactivated nuclei synthesize some transcripts not normally produced by the isolated nuclei and we have shown that a proportion of these possess amino acid-accepting activity. The significance of these results is discussed in relation to the control of gene activity in these cells.
Some restriction endonucleases generate a single-stranded nick at their recognition sequences in the presence of ethidium bromide (EtBr). This nick can then be extended to a single-stranded gap in which mutations can be introduced by a variety of techniques. To date, the templates used in these studies have largely contained a single recognition site for a given enzyme. Therefore, we have extended these studies to twelve enzymes for which multiple recognition sites exist in the template and show that, under appropriate conditions, one single-stranded nick is introduced per plasmid molecule.
Developments in understanding the pathophysiology of urinary incontinence caused by urethral sphincter mechanism incompetence (SMI) are reviewed. SMI is a multifactorial condition; affected bitches have shorter urethras, reduced urethral tone and more caudally positioned bladder necks than continent bitches. Other factors thought to be important in the aetiology of SMI include neutering, breed, body size, docking and obesity. Most of the recent developments in the understanding of SMI have resulted from radiographic and urodynamic investigations of normal and affected bitches. The urodynamic technique of urethral pressure profilometry is discussed and its contribution as an investigative tool reviewed.
The aim of this study was to document what changes in the resting and stressed urethral pressure profile occur in the incontinent bitch with urethral sphincter mechanism incompetence (SMI) immediately after colposuspension. Resting and stressed subtracted simultaneous urethral pressure profilometry was performed immediately pre- and postcolposuspension in 26 bitches diagnosed with SMI. All of the urethral pressure profiles were measured in anaesthetized bitches using a standard technique and two orientations of the catheter transducers (dorsal and left). Readable pre- and postoperative urethral pressure profiles were obtained in 20 of the 26 bitches. Subjective and objective evaluation of the profiles showed significant differences in the profiles pre- and postcolposuspension. Immediately postoperatively there were significant (P < .05) increases in functional profile length, maximum urethral closure pressure (MUCP), and distance between the bladder neck and the first negative respiratory peak and stressed spike. There was a significant (P < .001) decrease in the percentage of negative spikes extending below the resting intravesical pressure on the subtracted profile. The pressure transmission profiles were significantly (P < .001) altered by surgery. The findings presented support the hypothesis that colposuspension may restore continence by increasing pressure transmission to the proximal urethra and bladder neck. The results also suggest that immediately after surgery functional urethral length and urethral resistance are increased.
Vaginourethroplasty was performed in six bitches with infiltrative, obstructive urethral disease. The initial clinical signs included dysuria, hematuria, pollakiuria, and stranguria. Diagnosis was based on the clinical signs, plain radiography and contrast vagino-urethrogram examination, and urethral biopsy. Five dogs were diagnosed as having urethral neoplasia; the sixth dog had granulomatous urethritis. In all six dogs, up to 50% of the urethra and the urethral tubercle were resected to ensure adequate surgical margins. One dog was euthanatized 4 months after surgery for recurrence of a transitional cell carcinoma at the surgical site. Long-term resolution of the problem (minimum of 12 months) was achieved in the other five dogs. The frequency of serious complications, including urinary incontinence and ascending lower urinary tract infections, was low.
To evaluate ultrasonography as an alternative to contrast radiography for diagnosis of ectopic ureter in dogs, ultrasonography of the urinary tract was performed prospectively in a series of urinary incontinent dogs anesthetized for contrast radiography. Fourteen dogs had ectopic ureter based on surgical, necropsy or unequivocal contrast radiographic findings. There were eight females and six males of a variety of breeds; five were Labrador retrievers. Mean (range) age at the time of diagnosis was 1.2 (0.2-4) years for females and 3.5 (0.3-5) for males (p < 0.05). Ectopic ureters were unilateral in five dogs (2 left; 3 right) and bilateral in nine dogs. Both ultrasound images and contrast radiographs were positive for 21 (91%) ectopic ureters; the same two ectopic ureters were not detected using either modality. The termination of each of the five normal ureters was visible on ultrasound images; two (40%) were visible on radiographs. Other ultrasonographic findings included dilatation of the ectopic ureter and/or ipsilateral renal pelvis in ten (43%) instances, evidence of pyelonephritis in two dogs (with enlargement of the contralateral kidney in one dog), and urethral diverticuli in one dog. Ultrasonography is a practical diagnostic test for ectopic ureter in dogs. In this series there was close correlation between the ultrasonographic and contrast radiographic findings for each ectopic ureter, but ultrasonography enabled more accurate determination of normal ureteral anatomy.