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UNLABELLED: Lack of estrogen affects the urinary tract mainly by diminishing vascular, muscular and epithelial trophism, resulting in negative effects on continence in postmenopausal women. Therefore, the use of estrogens in these patients may revert these alterations and lead to an expressive improvement of the urinary symptoms. OBJECTIVE: Study the effect of topical estrogen therapy (conjugated equine estrogens, estriol or promestriene) in periurethral vessels detected by Dopplervelocimetric analysis using, as parameters: the number of vessels, resistance and pulsatility indexes, as well as the minimum diastolic value. METHODS: Forty-one postmenopausal women with stress urinary incontinence were randomized into three groups according to different types of topical estrogen received during 3 months. Group 1 received conjugated equine estrogens, group 2 received estriol and group 3 received promestriene. Periurethral Dopplervelocimetry analysis was done before estrogen administration and during treatment in all groups. RESULTS: We observed an increase in the number of the periurethral vessels in group 1 and group 2, being higher in group 1 than in group 2. The pulsatility index remained unchanged in all three groups. The resistance index at the periurethral vessels reduced only at the conjugated estrogen group (group 1). In this same group we noticed an increase in the mean minimal diastolic value, meaning a better periurethral vascularization. CONCLUSION: Topical conjugated equine estrogens and estriol were effective in increasing the number of periurethral vessels in postmenopausal women with urinary stress incontinence, with the conjugated equine estrogens being the most effective intervention studied.

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Our objective was to evaluate urodynamic and ultrasonographic findings after continence surgery. The study consisted of three groups of women according to the surgery performed: group I (Burch colposuspension) with 12 patients; group II (Kelly-Kennedy plication) with 10 patients; and group III (Gittes surgery) with 9 patients. Urodynamic study was done preoperatively and after surgery (on the 7th and 30th postoperative days, and at least 6 months after surgery) and ultrasonography of the bladder neck was performed to evaluate its position in relation to the inferior edge of the pubic symphysis and its mobility, both preoperatively and after surgery (30th postoperative day). All patients remained continent. We observed an increase in the first desire to void and maximum cystometric capacity after 6 months in groups I and II, respectively. There was no change in the urethral closure pressure profile in the three groups. Elevation of the bladder neck and decrease of its mobility were found by ultrasonography. Urinary continence after surgery is not the result of alterations in the urethral closure pressure profile, but rather of an elevation in the bladder neck and limitation of its mobility, which probably improves the abdominal pressure transmission rate to the proximal urethra.

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PURPOSE: While transformation of epithelial cells to a motile form is the first step in wound healing of the corneal epithelium, the migratory mechanism in these cells is not fully understood. We studied the expression of proto-oncogene mRNAs: c-fos; c-jun; fos B; jun B; jun D in injured corneal epithelium using in situ hybridization. Moreover, we examined immunolocalization of c-Fos and c-Jun protein products to elucidate the transcriptional activation prior to the onset of migration in corneal epithelium. METHODS: An epithelial defect was made on one cornea of 60 Wistar rats. The affected eye was enucleated immediately (within 5 min) or was allowed to heal for 15, 30, 60, 90, 120 and 180 min. Frozen sections were processed for in situ hybridization with c-fos, c-jun, fos B, jun B and jun D mRNAs or were stained with anti-c-fos and anti-c-jun antibodies. RESULTS: Fifteen min after the epithelial ablation, weak signals for c-fos and c-jun mRNAs were detected in the corneal epithelium surrounding the wound. These signals reached a peak 30 to 60 min after ablation, but were no longer evident at 120 min. Immunoreactivities for these proteins were also detected in the same area at 60 to 120 min after the epithelial ablation. Fos B mRNA was detected in the same region at 30 min after the ablation, and reached its peak after 30 to 60 min, but was no longer evident at 120 min. Jun B mRNA was detected in the epithelium around the defect 60 min after the ablation, later than the other proto-oncogenes, and reached its peak after 90 min. The message for jun D was detected in normal epithelium, and was not affected by wounding. CONCLUSIONS: These findings indicate that transcriptional activation of epithelial cells is initiated in the early phase after epithelial ablation, before the cells start to migrate, and that these proto-oncogene products may play important roles in wound healing in corneal epithelium. The time lag of the peak of expression of these proto-oncogenes in this process.

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A second-generation method of genotyping hepatitis C virus (HCV) was developed by the polymerase chain reaction (PCR) with sense as well as antisense primers deduced from the core gene. HCV RNA specimens extracted from sera were reverse-transcribed and amplified with universal primers in the first round of PCR to obtain fragments of 433 base pairs representing nucleotides 319-751. In the second round of PCR, portions of PCR products were amplified separately with sense and antisense primers specific for each of the five common genotypes prevailing across the world, i.e., I/1a, II/1b, III/2a, IV/2b and V/3a. The specificity of the method was verified by a panel of 177 HCV isolates of various genotypes in the genetic groups 1-9. It allowed clear differentiation of genotype I/1a from II/1b which was not always accomplished by the previous method. When 501 sera from blood donors and hepatitis patients with HCV viremia from various countries were genotyped by the second-generation method, 478 (95.4%) were classified into the five genotypes. HCV RNA samples from 23 (4.6%) sera were not classifiable into any of the five common genotypes and, by sequence analysis, 22 were found to be of four genotypes in group 4 and one of genotype 1c in Simmond's classification.

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OBJETIVO. Propoe-se, neste trabalho, analisar os resultados pós-operatórios de 367 cirurgias para correçao de incontinência urinária de esforço, quanto à época de aparecimento e ocorrência de recidivas. MATERIAL E MÉTODO. Os casos foram reunidos em quatro grupos: A = 206 (56,1 por cento) casos - cirurgia de Kelly-Kennedy; B = 37 (10,1 por cento) - cirurgia de Kelly- Kennedy associada à histerectomia vaginal; C = 95 (25,9 por cento) - cirurgia de Burch e, finalmente, D = 29 (7,9 por cento) - cirurgia de Burch associada à histerectomia total abdominal. RESULTADOS. O tempo de seguimento pós-operatório variou de 1 a 194 meses, com média de 31 meses. Observou-se incidência de recidivas de 20,1 por cento no grupo D e de 10,8 por cento no grupo B. Em 68,7 por cento das cirurgias por via vaginal e em 82,3 por cento pela abdominal, as recidivas ocorreram em menos de três anos. CONCLUSAO. Nao houve diferença estatisticamente significante no aparecimento de recidivas pós-operatórias nos três grupos estudados. A maior parte das recidivas ocorreu nos primeiros três anos após a cirurgia.

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BACKGROUND: The postoperative results of 367 surgical interventions for the correction of urinary stress incontinence were analyzed in terms of time of onset and occurrence of relapses. MATERIAL AND METHOD: The cases were assigned to four groups: A = 206 cases of Kelly-Kennedy surgery (56.1%); B = 37 cases of Kelly-Kennedy surgery associated with vaginal hysterectomy (10.1%); C = 95 cases of Burch surgery (25.9%), and finally, D = 29 cases of Burch surgery associated with total abdominal hysterectomy (7.9%). RESULTS: Postoperative follow-up ranged from 1 to 194 months, with a mean of 31 months. The incidence of relapses was 20.7% in group D, and 10.8% in group B. In 68.7% of the surgical interventions by vaginal route and in 82.3% of interventions by abdominal route, the relapses occurred within less than three years. CONCLUSION: The results were not different in the groups. The relapses occurred within less than three years in most cases.

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BACKGROUND: Minoxidil is an inhibitor of lysyl hydroxylase, an enzyme involved in collagen production, and decreases collagen production in vitro. We investigated the in vitro effects of minoxidil on behavior such as proliferation and migration of rabbit subconjunctival fibroblasts (SCFs). The ultrastructural effect of the drug on SCFs was also examined. METHODS: Proliferation of SCFs and closure of the defect produced in monolayer cultures in the presence or absence of minoxidil was studied. The ultrastructure of SCFs treated with minoxidil was also examined. RESULTS: Minoxidil inhibited SCF proliferation and the closure of the defect produced in monolayer cell sheets. Ultrastructural observations revealed extensive areas of irregularly dilated endoplasmic reticulum in cells treated with minoxidil, indicating the accumulation of protein, probably underhydroxylated collagen precursors, in the cisternae of endoplasmic reticulum. CONCLUSIONS: The results indicated that minoxidil attenuated cellular activities of SCFs such as proliferation and migration in vitro. The exact mechanism of the inhibitory effects of minoxidil on these cellular activities is unknown. The findings suggest that the drug might help to prevent bleb scarring after glaucoma filtering surgery.

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To investigate the changes in the corneal epithelial basement membrane following an alkali burn, we examined the immunolocalization of type IV collagen and laminin in the eye of the guinea pig burned with alkali. The burn damaged the corneal, limbal and conjunctival epithelium. After regeneration, basement membrane was interrupted, as indicated by laminin immunoreactivity. Type IV collagen immunoreactivity was transiently expressed in the early healing phase in the epithelial derived from both the cornea and conjunctiva, but was not seen in the normal corneal epithelial basement membrane. Later in the healing process, following transdifferentiation of the conjunctival epithelium into a cornea-like epithelium, its type IV collagen immunoreactivity was weaker than that in the basement membrane of the nontransdifferentiated epithelium. Conjunctival transdifferentiation during healing may have led to transient development of type IV collagen immunoreactivity.

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BACKGROUND: The spreading of epithelium is critical in the healing of corneal wounds. Such epithelial spreading requires the continuous production of protein and glycoprotein. To determine whether collagen production is required for the spreading of corneal epithelium, we studied the effects of inhibitors for collagen production on spreading of corneal epithelium in vitro. METHODS: We examined the effect of two proline analogs, L-azetidine 2-carboxylic acid and cis-hydroxyproline, a prolyl hydroxylase inhibitor, ethyl-3,4-dihydroxybenzoate, and a lysyl hydroxylase inhibitor, minoxidil, on the spreading of epithelium of organ-cultured rabbit cornea. RESULTS: Both analogs and inhibitors inhibited epithelial spreading in a dose-dependent manner. CONCLUSION: These observations indicate that collagen production may be involved in the spreading of corneal epithelium.

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We investigated the origin of fibronectin (FN) on five posterior and four anterior chamber explanted intraocular lenses (IOLs) using immunohistochemical methods. Cellular deposits (assumed to be macrophages) and fibrous or membrane-like proteinaceous deposits on the IOLs showed immunoreactivity to an antibody against cellular FN. These proteinaceous deposits were believed to be products of the cells that adhered to the IOLs.

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We investigated the altered immunolocalization of the components of epithelial basement membrane (BM) in rats with alkali burns of the cornea. Type IV collagen, laminin, and fibronectin were immunohistochemically stained in the alkali-burned corneas after various intervals. Linear laminin immunoreactivity, which represented the localization of the BM, was interrupted, probably by epithelial enzymes, under the regenerating epithelium after 24 h. A normal configuration was restored on day 7, when the central corneal BM showed a type IV collagen immunoreactivity that was not detected in BM of normal cornea. The exact cause of the development of type IV collagen immunoreactivity in the healing BM was not determined. Laminin, type IV collagen, and fibronectin, which were not detected in normal stroma, were all detected in the healing corneal stroma with repopulated keratocytes. Moreover, the BM zone under the regenerating epithelium showed fibronectin immunoreactivity on days 5-14. Distribution of these components of BM seemed to be normal 2 months postburn. The alteration of BM components is thought to be an important marker of the healing process in corneas burned with alkali.

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Recruitment of keratocytes into injured corneal stroma, and secretion of proteins including collagen in the cells are essential for wound healing of the corneal stroma. We examined the effect of a proline analog, cis-hydroxyproline, on the adhesion, migration and growth of rabbit keratocytes in vitro. This agent decreased the plating efficiency, migration and growth of the keratocytes in a dose-dependent manner. Reduction in these cellular activities may reflect altered functions of pericellular proteins such as collagen. Further studies are needed to determine which specific protein is involved.

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The gene of serine hydroxymethyltransferase (SHMT) of a thermophilic bacterium Bacillus stearothermophilus was expressed in Escherichia coli, and SHMT was successfully purified from the crude extract of E. coli in two steps while maintaining the enzymatic activity. The purification steps involved ammonium sulphate precipitation followed by high-performance liquid chromatographic separation using the anion-exchange column Fractogel EMD DEAE-650(S). In addition to the DEAE column, three other types of anion- and cation-exchange columns were also studied for their ability to separate SHMT, and the performance of the four columns were compared.

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Antibacterial activity of tinidazole (1-2-(ethylsulfonyl)-2-methyl-5-nitroimidazole) against anaerobic bacteria including Peptococcus, Peptostreptococcus, Eubacterium, Propionibacterium, Bacteroides and Fusobacterium was studied by agar dilution method comparing with metronidazole. In addition to this work, bactericidal effect of tinidazole and metronidazole against P. prevotii, B. fragilis ss. fragilis and F. varium was examined by quantitative culture method after incubation in GAM broth containing of 4 MIC, 2 MIC, 1 MIC and 1/2 MIC of both drugs against each of three strains for 1, 3, 6, 12 and 24 hours. All the strains of Peptococcus and Peptostreptococcus including P. anaerobius, P. saccharolyticus, P. prevotii and Ps. anaerobius and others were susceptible to a concentration of 6.25 mcg/ml of this drug. A concentration of 3.13 mcg/ml inhibited all strains of Bacteroides including B. fragilis ss. fragilis (12 strains), ss. vulgatus (5 strains), ss. thetaiotaomicron (4 strains) and ss. distasonis (2 strains). To this concentration all strains of Fusobacterium including F. varium (20 strains), F. mortiferum (2 strains) and other Fusobacterium sp. (5 strains) were susceptible. On the contrary, Propionibacterium acnes (6 strains) was resistant to 100 mcg/ml or more of tinidazole and metronidazole. The antibacterial activity of tinidazole was stronger against Bacteroides than that of metronidazole, while almost equal against Peptococcus, Peptostreptococcus, Eubacterium and Fusobacterium. Tinidazole was bactericidal against F. varium in a concentration of 2 MIC till 24 hours of incubation but did not show such an activity on B. fragilis ss. fragilis in same concentration even after 12 hours of incubation. On the other hand, metronidazole was bactericidal against B. fragilis ss. fragilis while was not against F. varium. Against P. prevotii bactericidal activity of both drugs was similar. Tinidazole as well as metronidazole is an excellent chemotherapeutic agent against anaerobic bacteria excluding Propionibacterium acnes and Bifidobacterium adolescentis.

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