A new measure of sexual function in women with pelvic floor disorders (PFD): the Pelvic Organ Prolapse/Incontinence Sexual Questionnaire, IUGA-Revised (PISQ-IR).

INTRODUCTION AND HYPOTHESIS: The objective of this study was to create a valid, reliable, and responsive sexual function measure in women with pelvic floor disorders (PFDs) for both sexually active (SA) and inactive (NSA) women. METHODS: Expert review identified concept gaps and generated items evaluated with cognitive interviews. Women underwent Pelvic Organ Prolapse Quantification (POPQ) exams and completed the Incontinence Severity Index (ISI), a prolapse question from the Epidemiology of Prolapse and Incontinence Questionnaire (ISI scores), the Pelvic Floor Distress Inventory-20 (PFDI-20), and the Female Sexual Function Index (FSFI). Principle components and orthogonal varimax rotation and principle factor analysis with oblique rotation identified item grouping. Cronbach's alpha measured internal consistency. Factor correlations evaluated criterion validation. Change scores compared to change scores in other measures evaluated responsiveness among women who underwent surgery. RESULTS: A total of 589 women gave baseline data, 200 returned surveys after treatment, and 147 provided test-retest data. For SA women, 3 subscales each in 2 domains (21 items) and for NSA women 2 subscales in each of 2 domains (12 items) emerged with robust psychometric properties. Cronbach's alpha ranged from .63 to .91. For SA women, correlations were in the anticipated direction with PFDI-20, ISI, and FSFI scores, POPQ, and EPIQ question #35 (all p < .05). PFDI-20, ISI, and FSFI subscale change scores correlated with Pelvic Organ Prolapse/Urinary Incontinence Sexual Questionnaire International Urogynecological Association-revised (PISQ-IR) factor change scores and with mean change scores in women who underwent surgery (all p < .05). For NSA women, PISQ-IR scores correlated with PFDI-20, ISI scores, and with EPIQ question #35 (all p < .05). No items demonstrated differences between test and retest (all p ≥ .05), indicating stability over time. CONCLUSIONS: The PISQ-IR is a valid, reliable, and responsive measure of sexual function.

Vitreous opacities affect scanning laser polarimetry measurements.

PURPOSE: To assess the effect of vitreous opacities on retinal nerve fiber layer retardation measurements obtained during scanning laser polarimetry. METHODS: Scanning laser polarimetry was performed in two eyes of two patients with vitreous opacities. RESULTS: The presence of a vitreous opacity within the measurement ellipse during scanning caused a marked, localized increase in polarization in the area of the opacity. This falsely increased the value obtained for the mean retinal nerve fiber layer thickness. Retinal nerve fiber layer thickness values were reduced when the opacity was not incorporated into the measurement ellipse. CONCLUSION: Artifact introduced by the presence of vitreous opacities can affect scanning laser polarimetry measurement reliability.

Assessment of retinal nerve fiber layer internal reflectivity in eyes with and without glaucoma using optical coherence tomography.

OBJECTIVE: To assess the internal reflectivity of the retinal nerve fiber layer in normal, ocular hypertensive, and glaucomatous eyes using optical coherence tomography. METHODS: All patients underwent complete ophthalmic examination and achromatic automated perimetry. Intraocular pressure was 21 mm Hg or less for low-tension glaucoma and normal eyes and at least 25 mm Hg on 2 separate occasions in ocular hypertensive and high-tension glaucoma eyes. All glaucomatous eyes had characteristic glaucomatous optic neuropathy and associated achromatic automated perimetry defect. Relative retinal nerve fiber layer internal reflectivity was measured on optical coherence tomography images using a software program of our own design. RESULTS: We enrolled 98 eyes (19 normal, 34 ocular hypertensive, 17 high-tension glaucoma, and 28 low-tension glaucoma). Relative internal reflectivity was less in eyes with glaucoma than in normal (P<.001, t test) and ocular hypertensive eyes (P<.001, t test). There was no difference in relative internal reflectivity between normal and ocular hypertensive eyes (P =.32) and between eyes with high-tension glaucoma and low-tension glaucoma (P =.43). Internal reflectivity correlated with mean deviation on achromated automatic perimetry (r(2) = 0.49, P<.001, quadratic regression analysis). CONCLUSION: Relative retinal nerve fiber layer internal reflectivity may provide useful information about the extent of retinal nerve fiber layer injury in glaucoma. Arch Ophthalmol. 2000;118:1044-1047

Scanning laser polarimetry measurements after laser-assisted in situ keratomileusis.

PURPOSE: To evaluate the effect of laser-assisted in situ keratomileusis on retinal nerve fiber layer thickness measurements obtained with scanning laser polarimetry. METHODS: Thirteen consecutive eyes (13 patients) undergoing laser-assisted in situ keratomileusis were enrolled in this prospective study. Scanning laser polarimetry (NFA-GDx; Laser Diagnostic Technologies, Inc, San Diego, California) examination was performed 1 week before and 1 to 8 weeks after laser-assisted in situ keratomileusis surgery. Intraocular pressure was normal at all preoperative and postoperative examinations. Total mean, and superior, temporal, inferior, and nasal mean retinal nerve fiber layer thickness values before and after laser-assisted in situ keratomileusis were compared by Student paired t test. RESULTS: Mean +/- SD patient age was 34.6 +/- 10.9 years (range, 20 to 56 years). Mean +/- SD preoperative spherical equivalent refractive error was -6.6 +/- 3.1 diopters (range, -3.25 to -13.25 diopters) and mean +/- SD spherical equivalent refractive surgical correction was -6.2 +/- 3.0 diopters (range, -2.9 to -12.25 diopters). Total mean retinal nerve fiber layer and superior, inferior, temporal, and nasal mean retinal nerve fiber layer thicknesses were thinner after laser-assisted in situ keratomileusis (P =.01, for all comparisons, paired t test). CONCLUSIONS: Measurements of the retinal nerve fiber layer with scanning laser polarimetry depend on a corneal compensator inherent in the device. Keratorefractive surgery may affect scanning laser polarimetry measurements.

Heidelberg retina tomography and optical coherence tomography in normal, ocular-hypertensive, and glaucomatous eyes.

PURPOSE: To evaluate optic disc and retinal nerve fiber layer (RNFL) appearance in normal, ocular-hypertensive, and glaucomatous eyes undergoing confocal scanning laser ophthalmoscopy and optical coherence tomography (OCT). DESIGN: Prospective, cross-sectional study. PARTICIPANTS: Seventy-eight eyes of 78 consecutive normal (n = 17), ocular-hypertensive (n = 23), and glaucomatous subjects (n = 38) were enrolled. METHODS: Each patient underwent complete ophthalmic examination, achromatic automated perimetry, confocal scanning laser ophthalmoscopy (Heidelberg Retinal Tomography [HRT]), and OCT. Topographic HRT parameters (disc area, cup-disc ratio, rim area, rim volume, cup shape measure, mean RNFL thickness, and cross-sectional area) and mean OCT-generated RNFL thickness were evaluated in each group. MAIN OUTCOME MEASURES: OCT and HRT assessment of optic disc and RNFL anatomy. RESULTS: OCT RNFL thickness showed no difference between normal and ocular-hypertensive eyes (P = 0.15) but was significantly less in glaucomatous eyes (P < 0.001). HRT measurements of rim area, cup-disc ratio, cup shape measure, RNFL thickness, and RNFL cross-sectional area were significantly less in glaucomatous eyes (all P < 0.005) and were correlated with mean OCT RNFL thickness (all P < 0.02). RNFL thickness using OCT or HRT was highly correlated with visual field mean defect during achromatic perimetry (P < 0.0001). CONCLUSION: Both HRT and OCT can differentiate glaucomatous from nonglaucomatous eyes. RNFL thickness measurements using OCT correspond to disc topographic parameters using HRT.

Streptococcus pneumoniae DNA polymerase I lacks 3'-to-5' exonuclease activity: localization of the 5'-to-3' exonucleolytic domain.

The Streptococcus pneumoniae polA gene was altered at various positions by deletions and insertions. The polypeptides encoded by these mutant polA genes were identified in S. pneumoniae. Three of them were enzymatically active. One was a fused protein containing the first 11 amino acid residues of gene 10 from coliphage T7 and the carboxyl-terminal two-thirds of pneumococcal DNA polymerase I; it possessed only polymerase activity. The other two enzymatically active proteins, which contained 620 and 351 amino acid residues from the amino terminus, respectively, lacked polymerase activity and showed only exonuclease activity. These two polymerase-deficient proteins and the wild-type protein were hyperproduced in Escherichia coli and purified. In contrast to the DNA polymerase I of Escherichia coli but similar to the corresponding enzyme of Thermus aquaticus, the pneumococcal enzyme appeared to lack 3'-to-5' exonuclease activity. The 5'-to-3' exonuclease domain was located in the amino-terminal region of the wild-type pneumococcal protein. This exonuclease activity excised deoxyribonucleoside 5'-monophosphate from both double- and single-stranded DNAs. It degraded oligonucleotide substrates to a decameric final product.

The polymerase domain of Streptococcus pneumoniae DNA polymerase I. High expression, purification and characterization.

The 3'-terminal two-thirds of the Streptococcus pneumoniae polA gene was cloned in an Escherichia coli genefusion vector with inducible expression. The resulting recombinant plasmid (pSM10) directs the hyperproduction of a polypeptide of 70.6 kDa corresponding to the C-terminal fragment of pneumococcal DNA polymerase I. Induced cells synthesized catalytically active protein to the extent of 7% of the total soluble protein in the cells. The polymerase fragment was purified to greater than 90% homogeneity with a yield of 1.5 mg pure protein/l culture. The protein has DNA polymerase activity, but no exonuclease activity. The enzyme requires a divalent cation (MgCl2 or MnCl2) for polymerization of DNA. Comparison of the mutant and wild-type pneumococcal polymerases shows that the construction did not affect the enzymatic affinity for the various substrates. The mutant protein, like its parent DNA polymerase I, exhibited an intermediate level of activity with primed single-stranded DNA. At high molar ratio of enzyme/DNA substrate, the polymerase fragment catalyzes strand displacement and switching after completing the replication of a primed single-stranded M13 DNA molecule.