Role of fibulin-5 insufficiency and prolapse progression on murine vaginal biomechanical function.

The vagina plays a critical role in supporting the pelvic organs and loss of support leads to pelvic organ prolapse. It is unknown what microstructural changes influence prolapse progression nor how decreased elastic fibers contributes to vaginal remodeling and smooth muscle contractility. The objective for this study was to evaluate the effect of fibulin-5 haploinsufficiency, and deficiency with progressive prolapse on the biaxial contractile and biomechanical function of the murine vagina. Vaginas from wildtype (n = 13), haploinsufficient (n = 13), and deficient mice with grade 1 (n = 9) and grade 2 or 3 (n = 9) prolapse were explanted for biaxial contractile and biomechanical testing. Multiaxial histology (n = 3/group) evaluated elastic and collagen fiber microstructure. Western blotting quantified protein expression (n = 6/group). A one-way ANOVA or Kruskal-Wallis test evaluated statistical significance. Pearson's or Spearman's test determined correlations with prolapse grade. Axial contractility decreased with fibulin-5 deficiency and POP (p < 0.001), negatively correlated with prolapse grade (ρ = - 0.80; p < 0.001), and positively correlated with muscularis elastin area fraction (ρ = - 0.78; p = 0.004). Circumferential (ρ = 0.71; p < 0.001) and axial (ρ = 0.69; p < 0.001) vaginal wall stresses positively correlated with prolapse grade. These findings demonstrated that fibulin-5 deficiency and prolapse progression decreased vaginal contractility and increased vaginal wall stress. Future work is needed to better understand the processes that contribute to prolapse progression in order to guide diagnostic, preventative, and treatment strategies.

Metastatic Acinic Cell Carcinoma to the Vagina: A First Reported Case.

This is a case of a 62-year-old woman with a remote history of acinic cell carcinoma of the parotid gland, who presented with a palpable vaginal mass, anterior vaginal wall prolapse, and stress urinary incontinence. A 2 cm firm mobile mass on the anterior vaginal wall was found on clinical examination. A computed tomographic scan revealed a mass between the vaginal vault and bladder that was eventually surgically excised. The histology, supported by the immunohistochemistry, revealed metastatic acinic cell carcinoma to the vagina after 37 years of her initial diagnosis. This is the first reported case in the literature to occur in the vagina.

[Outcomes of ER:YAG LASER treatment of stress urinary incontinence in women].

AIM: To evaluate the effectiveness of a new method of treatment for stress urinary incontinence in women using an ER: YAG laser in SMOOTH mode and investigate pathophysiological and pathomorphological changes induced by erbium laser. MATERIALS AND METHODS: This study comprised 98 women aged 37-63 years, who between 2014 and 2016 were diagnosed with SUI (type 1 and 2a, 2b) and grade 0-2 vaginal prolapse. The treatment was performed with a 2940 nm Er:YAG laser (Fotona, Slovenia) using a SMOOTH mode. Clinical assessment included PFIQ-7 and PISQ-12 questionnaires, uroflowmetry, laser Doppler flowmetry and biopsy of the anterior vaginal wall. The examination was carried out at baseline and 1-2 months after the treatment. RESULTS: The effectiveness of treatment was 73%. There was no deterioration after the procedure. Analysis of PFIQ-7 and PISQ-12 questionnaires showed that patients with mild incontinence had the greatest difference between pre- and posttreatment results. Uroflowmetry parameters improved in a majority of patients. Results of laser Doppler flowmetry demonstrated the improvement of blood flow in the microvascular bed. An important feature of the vaginal biopsy after laser exposure was an increase in neoangiogenesis. DISCUSSION: The findings of questionnaires and clinical evaluation of patients with SUI and vaginal prolapse before and after treatment with Er: YAG laser showed high therapeutic effectiveness of this treatment modality. CONCLUSION: Clinical effectiveness of ER: YAG laser in SMOOTH mode was 73%. Patients with type 1 and 2a SUI and mild or moderate incontinence have the best prognosis after treatment with this method.

Effects of Pneumoperitoneum and the Steep Trendelenburg Position on Heart Rate Variability and Cerebral Oxygenation during Robotic Sacrocolpopexy.

STUDY OBJECTIVE: The aim of this study was to investigate how steep Trendelenburg positioning with pneumoperitoneum modifies brain oxygenation and autonomic nervous system modulation of heart rate variability during robotic sacrocolpopexy. DESIGN: Prospective study (Canadian Task Force classification III). SETTING: Rambam Health Care Campus. PATIENTS: Eighteen women who underwent robotic sacrocolpopexy for treatment of uterovaginal or vaginal apical prolapse. INTERVENTIONS: Robotic sacrocolpopexy. MEASUREMENTS AND MAIN RESULTS: A 5-minute computerized electrocardiogram, cerebral O2 saturation (cSO2), systemic O2 saturation, heart rate (HR), diastolic blood pressure (BP), systolic BP, and end-tidal CO2 tension were recorded immediately after anesthesia induction (baseline phase) and after alterations in positioning and in intra-abdominal pressure. HR variability was assessed in time and frequency domains. Cerebral oxygenation was measured by the technology of near-infrared spectrometry. cSO2 at baseline was 73% ± 9%, with minor and insignificant elevation during the operation. Mean HR decreased significantly when the steep Trendelenburg position was implemented (66 ± 10 vs 55 ± 9 bpm, p < .05) and returned gradually to baseline with advancement of the operation and the decrease in intra-abdominal pressure. Concomitant with this decrease, the power of both arms of the autonomic nervous system increased significantly (2.8 ± .8 vs 3.3 ± .9 ms2/Hz and 2.5 ± 1.2 vs 3.2 ± .9 ms2/Hz, respectively, p < .05). All these effects occurred without any significant shifts in systolic or diastolic BP or in systemic or cerebral oxygenation. CONCLUSION: This study supports the safety of robotic sacrocolpopexy performed with steep Trendelenburg positioning with pneumoperitoneum. Only minor alterations were observed in cerebral oxygenation and autonomic perturbations, which did not cause clinically significant alterations in HR rate and HR variability.

Hypoxia Induces Apoptosis through HIF-1α Signaling Pathway in Human Uterosacral Ligaments of Pelvic Organ Prolapse.

The purpose of this study is to evaluate the expression of hypoxia-inducible factor-1α (HIF-1α) in women uterosacral ligament tissues with pelvic organ prolapse and women with normal uterine support structures and illuminate its relationship with apoptosis. Samples were collected from 38 women with pelvic organ prolapse and 31 age matched controls. The expression levels of HIF-1α and BNIP3 in the uterosacral ligaments were measured using immunohistochemistry, qRT-PCR, and Western blot. To assess apoptosis we performed TUNEL assay and Western blot analyses. Lastly, the short form of the Pelvic Floor Impact Questionnaire-7 (PFIQ-7) was used to evaluate prognosis of surgical patients and twenty patients finished the follow-up. The expressions of HIF-1α and BNIP3 in the uterosacral ligaments were significantly higher in patients with pelvic organ prolapse than in control group. Pearson's correlation test revealed significant positive correlations between HIF-1α and apoptosis index. Similarly, Western blot analysis showed the expression of proapoptosis proteins (Bax and Bad), Cytochrome-c, cleaved caspase-3, and caspase-9 in patients with pelvic organ prolapse was upregulated. The PFIQ-7 scores were higher in HIF-1α positive group than in the negative group. Hypoxia may contribute to the pathological process of pelvic organ prolapse by increasing apoptosis via activating HIF-1α signaling pathway.

Estrogen receptor expression and vessel density in the vagina wall in postmenopausal women with prolapse.

After menopause, critically estrogen low levels result in modifications in vaginal wall. This cross-sectional study aims to determine whether there is a change in the number of vessels in the lamina propria of the vagina after menopause in parallel to the ER-alpha expression on the vaginal wall. Twelve women who underwent a genital surgery for genital prolapse up to grade II were selected. They were divided into two groups: a premenopausal group (PG) consisting of six women who were 18-40 years old with FSH levels =12 mIU/ml and regular cycles, and a menopausal group (MG) consisting of six women at least one year after menopause who were <65 years old with FSH levels =40 mIU/ml. Slides were stained for ER-alpha immunohistochemistry, and an endothelial cell marker CD3 was used to label vessels which were identified by using a system for morphometry. The number of vessels was significantly higher in the PG than in the MG both on the anterior wall (PG: 1.055 ± 145.8 vessels/mm(2), MG: 346.6 ± 209.9 vessels/mm(2), p<0.0001) and on the posterior wall (PG: 1064 ± 303.3 vessels/mm(2), MG: 348.6 ± 167.3 vessels/mm(2), p=0.0005). The ER-alpha score was significantly higher in the PG than the score for the MG on both the anterior and posterior walls (PG: 6.0 ± 0.52, MG: 2.5 ± 0.89, p=0.007; PG: 5.8 ± 0.79, MG: 2.7 ± 0.95, p=0.03, respectively). There was a positive correlation between the ER-alpha score and the vessel concentration on the anterior (r=0.6656, p=0.018) and posterior (r=0.6738, p=0.016) vaginal walls. Age was strongly negatively correlated with vessel concentration on the vaginal walls (respectively r=-0.9033, p<0.0001, r=-0.7440, p=0.0055). Therefore, postmenopausal women with genital prolapse have a smaller number of vessels on the vaginal wall compared to normoestrogenic controls with the same pathological condition. Hypoestrogenism and advancing age are factors that are associated to these changes.

The expression of glutathione peroxidase-1 and the anabolism of collagen regulation pathway transforming growth factor-beta1-connective tissue growth factor in women with uterine prolapse and the clinic significance.

OBJECTIVES: To investigate the expression of the anabolism of collagen regulation pathways connective tissue growth factor (CTGF) -transforming growth factor-betal (TGF-beta1) and glutathione peroxidase-1 (GPx1) in women with uterine prolapse and a study of the clinic significance. MATERIALS AND METHODS: The expression of TGF-beta1, CTGF, and GPx1 was detected by immunohistochemical staining in pubocervical fascia tissue of 30 women with uterine prolapse, including ten cases of POP-QII, ten cases of POP-QIII, ten cases of POP-QIV, and 20 cases were control group with non-prolapse and non-malignant lesions. RESULTS: There was a negative correlation between the POP-Q and expression of TGF-beta1. With the increase of POP-Q degree, the expression degree of TGF-beta1 decreased correspondingly, which also applied to CTGF and GPx1. On the other hand, there was a positive correlation between TGF-beta1 and CTGF. The synergistic change trend was found between TGF-beta1 and CTGF. It could also be seen between CTGF and GPx1 and betweenTGF-beta1 and GPx1. CONCLUSION: The expression of the antioxidase GPx1 in pelvic support structure of POP women was decreased, which resulted in the antioxidation reduced. It could break the balance of oxidation and antioxidation in pelvic support structure, and may induce an increase of ROS level and the down-regulation of TGF-beta1-CTGF pathway. It could inhibit the anabolism of collagen and injury the pelvic support structure, thus promoting the occurrence and development of POP.

Relationship between the expression of fibulin-3 and anterior vaginal wall prolapse.

Anterior vaginal wall prolapse is the most common type of pelvic organ prolapse. Vaginal wall samples were obtained from women with (n =12) and without (n =12) anterior vaginal wall prolapse. No reports have been published on the content of fibulin-3 in the vaginal walls of patients with prolapse; thus, we compared the expression of fibulin-3 in the vaginal walls of women with and without anterior vaginal wall prolapse. RT-PCR was performed to measure mRNA expression and the expression of protein was assessed by immunohistochemistry. Statistical analysis was performed to determine differences between the two groups of women. Age, parity and menopausal status did not differ between women with and without prolapse. The expressions of fibulin-3 mRNA and protein were not different between the prolapse and no prolapse groups. It is unlikely that abnormal expression of fibulin-3 has a major role in the pathogenesis of anterior vaginal wall prolapse.

Composição da matriz extracelular do ligamento cardinal de mulheres na pós-menopausa com e sem prolapso uterino / Composition of extracellular matrix of the cardinal ligament in postmenopausal women with and without uterine prolapse

O prolapso uterino tem sua incidência aumentada na pós-menopausa. O objetivo deste estudo é identificar as alterações na matriz extracelular do ligamento cardinal associadas à menopausa e ao prolapso uterino. Ligamento cardinal de três diferentes grupos de mulheres, pré-menopausa, prolapso uterino e pós-menopausa, foram identificados e biopsiados durante 57 histerectomias abdominais ou vaginais. As amostras foram processadas por métodos bioquímicos para caracterização e quantificação de glicosaminoglicanos sulfatados e colágeno. As concentrações relativas de glicosaminoglicanos foram obtidas por eletroforese. Procedimentos histológicos foram feitos para identificar fibras elásticas (Weigert), distribuição de colágeno (Picro Sirius) e decorin (imunohistoquímica). Nossos resultados mostraram aumento na concentração de GAG de 72,2%, redução na concentração de colágeno de 37% e diminuição de 22% de fibras elásticas no grupo de prolapso uterino quando comparado ao grupo da pós-menopausa (p<0,05, p<0,04 e p<0,05 respectivamente). As concentrações relativas de glicosaminoglicanos sulfatados para condroitin sulfato, heparan sulfato e dermatan sulfato não mostraram diferenças entre os três grupos. A organização do colágeno foi similar entre os três grupos e a marcação do decorin pareceu estar diminuída no grupo de prolapso uterino. Nossos resultados indicam alterações no metabolismo do tecido conjuntivo. O ligamento cardinal da mulher na pós-menopausa possui uma matriz extracelular mais densa. Esta alteração não ocorre na mulher com prolapso uterino.

Uterine prolapse has increase of incidence after menopause. The aim of this study was to identify the changes in extracellular matrix of cardinal ligaments associated to menopause and uterine prolapse. Cardinal ligament of 3 different groups (pre-menopause, menopause and uterine prolapse) are identified and biopsied during 57 women's abdominal or vaginal hysterectomy. Biopsy specimens were assessed by biochemical methods to characterize and quantify sulfated glycosaminoglycans and collagen. Relative concentrations of GAG were obtained by electrophoresis. Histological procedures are made to identify elastic fibers (Weigert) collagen distribution (Picro sirius) and decorin (immunohistochemistry). Our results showed increase in GAG concentration 72.2% in uterine prolapse group compared to menopause group (p<0.05). Collagen concentration was 37% lower in uterine prolapse group compared to menopause group (p<0.04). Relative concentration of GAG: heparan sulfate, chondroitin sulfate and dermatan sulfate showed no differences among three groups. Elastic fibers showed a significant reduction of aproximately 22% uterine prolapse group compared to menopause group (p<0.05). Collagen organization was similar in three groups and the staining pattern of decorin seemed to be decreased in uterine prolapse group. Our results indicate changes in connective tissue metabolism. Cardinal ligament in postmenopausal women has a denser extracellular matrix. This change is not observed in women with uterine prolapse.

Expression of matrix metalloproteinase-2 and tissue inhibitors of metalloproteinase-1 (TIMP-1, TIMP-2 and TIMP-3) in women with uterine prolapse but without urinary incontinence.

OBJECTIVE: To investigate the activities of matrix metalloproteinase-2 (MMP-2) and its inhibitors, tissue inhibitor of metalloproteinase-1, -2 and -3 (TIMP-1, TIMP-2 and TIMP-3), in the pelvic support and nonsupport tissue of women with uterine prolapse but without urinary incontinence. STUDY DESIGN: Paired samples of uterosacral ligament and cervical tissue were obtained from 11 postmenopausal and 8 premenopausal women with severe uterine prolapse. Nine premenopausal women without prolapse were selected as normal controls. Immunoreactivity of MMP-2 and TIMPs was demonstrated by immunohistochemistry. Steady state of MMP-2 as well as TIMPs messenger RNA (mRNA) expression was analyzed by polymerase chain reaction (PCR) with quantitative expression determined by multiplex PCR. RESULTS: A significantly higher expression of MMP-2 mRNA and lower expression of TIMP-2 mRNA were found in uterosacral ligament in uterine prolapse women compared to controls. In the cervical tissue, however, the MMP-2 and TIMPs mRNA expression was comparable between prolapse and control groups. With regard to menopausal status, there was no significant difference in MMP-2 and TIMPs mRNA expression between premenopausal and postmenopausal women with uterine prolapse. CONCLUSIONS: An increase in MMP-2 mRNA and a decrease in TIMP-2 mRNA expression in uterosacral ligament are related to uterine prolapse in women without urinary incontinence.

[Microarray analysis of gene expression profiles in pelvic organ prolapse].

OBJECTIVE: To identify the differentially expressed genes in cardinal ligament between patients with pelvic organ prolapse (POP) and postmenopausal women without POP by Human Genome Expression Chip and explore the potential molecular mechanism involved in POP. METHODS: From January to May, 2007, cardinal ligament samples were obtained from 3 postmenopausal patients with POP-Q stage III and 3 postmenopausal patients underwent hysterectomy due to other benign gynecologic diseases without POP in Peking Union Medical College Hospital. HE and Masson's trichrome staining was used to verify tissue origin and inspect histological changes. Those differentially expressed genes in cardinal ligaments were identified by Human Genome Chip and further interrogated with Gene Ontology (GO) and Pathway Analysis. Those remarkable expressed genes were confirmed by qRT-PCR. RESULTS: Alterations of ligament architecture in POP patients included disarrangement and collapse of smooth muscle bundles and collagen fibers. A total of 179 differentially expressed genes were screened between POP and non-POP cardinal ligament tissue, including 20 functional unknown genes. A total of 107 genes were upregulated in POP group, while 72 genes downregulated. Those differentially genes were revealed associated with multiple functional proteins and metabolic pathways by biological analysis. Among these, Wnt signaling pathway exhibited the most remarkable changes. Real-time quantitative PCR showed the genes of COL1A1, DKK1, SFRP1, FZD5, WNT16b in POP group (2.98+/-1.40, 3.03+/-0.48, 8.13+/-4.42, 5.19+/-3.50, 12.40+/-3.88) were upregulated significantly compared with non-POP group (1.09+/-0.08, 1.20+/-0.18, 0.41+/-0.51, 0.87+/-0.24, 1.40+/-0.47; P<0.05). CONCLUSIONS: The pathophysiology of POP is complex and associated with multiple functional proteins and metabolic pathways. Among these, the antagonist DKK1, SFRP1 in Wnt signaling pathway may contribute to a neurodegenerative role in POP development.

Differential expression of smooth muscle regulatory proteins in the uterosacral ligaments of women with uterine prolapse.

OBJECTIVE: To compare smooth muscle regulatory protein expression in the uterosacral ligament (USL) of women with and without uterine prolapse. STUDY DESIGN: USLs ligament were sampled in women with (n = 9) or without (n = 9) uterine prolapse. Caldesmon, smooth muscle actin (SMA), myosin heavy chain, and zinc finger protein messenger RNA expression was assessed by quantitative real-time polymerase chain reaction. Immunohistochemistry and digital image analysis were used to determine protein expression. RESULTS: Caldesmon messenger RNA expression and the ratio of caldesmon-SMA messenger RNA expression was significantly increased in the USL from women with uterine prolapse compared with women without prolapse (caldesmon mean +/- standard deviation messenger RNA, 0.81 +/- 0.46 vs 0.39 +/- 0.16; P = .01 and caldesmon-SMA messenger RNA ratio, mean +/- standard deviation, 0.11 +/- 0.04 vs 0.07 +/- 0.02; P = .01). In addition, the ratio of caldesmon-SMA staining was significantly increased in women with uterine prolapse compared with women without prolapse (mean +/- standard deviation, 0.44 +/- 0.28 vs 0.28 +/- 0.16; P = .03). CONCLUSION: Uterine prolapse is associated with an increased ratio of caldesmon-SMA actin expression.

Alteration of vaginal elastin metabolism in women with pelvic organ prolapse.

OBJECTIVE: To compare elastin metabolism in the vagina of women with and without pelvic organ prolapse and to define the regulation of this process by hormone therapy (HT). METHODS: Eighty-seven histologically confirmed full-thickness vaginal biopsies were procured from study participants at time of surgery. Premenopausal women with no prolapse served as controls. Women with prolapse were divided into three groups: premenopausal, postmenopausal not on HT, and postmenopausal on HT. The epithelium was excised leaving the subepithelium, muscularis, and adventitia for analyses. The elastin precursor, tropoelastin, was measured by immunoblotting and mature elastin protein via a desmosine cross-link radioimmunoassay. Matrix metalloproteinases (MMPs)-2 and -9 were quantitated by gelatin zymography. Data were analyzed using Kruskal-Wallis test and post hoc analysis using the Mann-Whitney U test. RESULTS: Tropoelastin (432%), mature elastin (55%), proMMP-9 (90%), and active MMP-9 (106%) were increased in women with prolapse relative to those in the control group while active MMP-2 (41%) was decreased. Comparison of tropoelastin and mature elastin values obtained from the same women showed them to be independently regulated (r=0.19). Interestingly, the highest amount of both proteins occurred in postmenopausal patients not on HT. CONCLUSION: Elastin metabolism is altered in the vagina of women with prolapse relative to those in the control group, suggesting that vaginal tissue is rapidly remodeling in response to mechanical stretch. We found that elastin levels are highest in the absence of hormones.

Differential expression of fibulins in the uterosacral ligaments of women with uterine prolapse.

PURPOSE: To compare fibulin-3 (FIB-3) and fibulin-5 (FIB-5) expressions in uterosacral ligaments (USL) of women with and without uterine prolapse. STUDY DESIGN: USL were sampled in a standardized fashion from women with (n = 8) or without (n = 8) uterine prolapse. Quantitative Real-Time PCR was performed to measure mRNA and immunohistochemistry to assess protein expression. RESULTS: FIB-3 mRNA expression and FIB-3 staining intensity was similar in the USL of women with and without uterine prolapse {[(FIB-3 mean +/- SD mRNA relative units) 0.45 +/- 0.41 vs. 0.46 +/- 0.82, NS] and [Intensity score, median (range), 1(0-1) vs. 1(0-1), NS]}. Both FIB-5 mRNA expression and FIB-5 staining intensity was significantly decreased in USL from women with uterine prolapse compared to women without prolapse {[(FIB-5 mean +/- SD mRNA relative units) 0.07 +/- 0.02 vs. 0.26 +/- 0.20, P = 0.02] and [Intensity score, median (range), 0(0-2) vs. 3(2-3), P = 0.002]}. CONCLUSION: FIB-5 expression is decreased in USL of women with uterine prolapse.

Extracellular matrix expression of human prolapsed vaginal wall.

OBJECTIVE: To compare the mRNA expression of extracellular matrix (ECM) proteins in postmenopausal prolapsed versus non-prolapsed anterior vaginal wall (AVW) tissue. We hypothesized that the weakening of the tissue leading to prolapse was due to decreased collagen production from a downregulation at the transcriptional level. METHODS: Following IRB approval, full thickness samples of redundant AVW were excised from consecutive age-equivalent, postmenopausal, women undergoing cystocele repair (prolapse, stage III or IV), or radical cystectomy (control, no clinical findings of prolapse). Total RNA was isolated, cDNA was synthesized, and quantitative real-time polymerase chain reaction (PCR) was conducted to assess the mRNA expression of collagens type I and III, pro-elastin, MMP3, MMP10, and MMP11. The significance of the difference of mRNA expression between prolapse and control tissues was tested using Student's t-test followed by Mann-Whitney Rank Sum Test. RESULTS: A 5.3-fold increase in collagen type I mRNA was found in prolapse (n = 47) over control (n = 7) tissues (P = 0.009). Type III collagen mRNA was also significantly increased to a 3.3 times higher level (P = 0.017). The ratio of type III to type I was decreased from 15.6 in controls to 9.7 in prolapse. An increasing trend in pro-elastin and MMP mRNA expression was found in prolapse, but this was not statistically significant. CONCLUSION: In this controlled study, the increase found in collagen mRNA expression disproved our hypothesis. To the contrary, this defective prolapsed tissue can signal its need for ECM replenishment. The message, however, is not being effectively translated to assist in tissue remodeling.

Matrix metalloproteinases-1, -2 expression in uterosacral ligaments from women with pelvic organ prolapse.

OBJECTIVE: This study investigated matrix metalloproteinase-1 (MMP-1) and matrix metalloproteinase-2 (MMP-2) immunohistochemical expression in uterosacral ligament biopsies from women with pelvic organ prolapse (POP), and controls with normal pelvic support. DESIGN: Prospective observational experimental study. SETTING: A tertiary Urogynecology Unit and Institute of Pathology, University Hospital Split, Croatia. POPULATION: Women referred for hysterectomy for prolapse or benign gynecological disease. METHODS: Eighty postmenopausal women were included in the study after Ethical Committee approval and informed consent. During surgery, uterosacral ligament biopsies were obtained from patients with POP (n=40) and women without evidence of pelvic floor weakening (n=40). Immunohistochemistry for MMP-1 and MMP-2 was performed on formaline fixed and paraffin embedded sections. Statistical evaluations were made by Student t-test or chi(2) test. MAIN OUTCOME MEASURES: Immunohistochemical expression of MMP-1 and MMP-2 in the uterosacral ligaments of women with and without genital prolapse. RESULTS: Forty women with POP and 40 controls without POP were included. The controls were matched to the women with POP in age, body mass index, parity and duration of postmenopausis. A significant increase in MMP-1 immunohistochemical expression was seen in uterosacral ligament tissue from women with POP (P=0.029). In contrast, there was no difference in immunohistochemical expression of MMP-2 between women with POP and those without (P=0.899). CONCLUSION: These data suggest that MMP-1 may be marker of collagen degradation. Increased MMP-1 immunohistochemical expression in uterosacral ligaments is associated with urogenital prolapse.

Caldesmon expression is decreased in women with anterior vaginal wall prolapse: a pilot study.

INTRODUCTION AND HYPOTHESIS: The purpose of this study is to compare vaginal caldesmon expression in women with and without anterior vaginal wall prolapse. METHODS: Vaginal tissues were sampled in women with (n = 11) or without (n = 11) vaginal wall prolapse. Caldesmon messenger RNA (mRNA) expression was assessed by quantitative real-time polymerase chain reaction. Immunohistochemistry and digital image analysis were used to determine caldesmon protein expression in the histologic sections. RESULTS: There were no significant differences in demographic data between the two groups. Caldesmon mRNA expression was significantly decreased in the vaginal tissue from women with anterior vaginal wall prolapse compared to women without prolapse [(caldesmon mean +/- SD mRNA expression in relative units) 0.03 +/- 0.03 vs 0.17 +/- 0.17, P = 0.02]. The fractional area of nonvascular caldesmon staining in the vagina of women with anterior vaginal wall prolapse was significantly decreased compared to women without prolapse [mean +/- SD (0.09 +/- 0.04 vs 0.16 +/- 0.09, P = 0.03)]. CONCLUSIONS: Vaginal caldesmon expression is significantly decreased in women with anterior vaginal wall prolapse compared to normal subjects.

Low copy number and high 4977 deletion of mitochondrial DNA in uterosacral ligaments are associated with pelvic organ prolapse progression.

INTRODUCTION AND HYPOTHESIS: The pathophysiology of pelvic organ prolapse (POP) is related to aging in the pelvic organ support, and mitochondrial dysfunction is one of the major contributors to aging. Therefore, the objective of this study was to investigate the correlation between alternations of mitochondrial DNA and progression of POP. METHODS: Polymerase chain reaction (PCR) was applied in the present study. Uterosacral ligaments (UL) were obtained from 45 POP patients and 38 myoma patients without POP. Chi-square test, Student's t-test, Mann-Whitney U test, and Spearman correlation analysis were applied in the comparison between POP and non-POP patients. RESULTS: The results revealed that significant depletion of mitochondrial DNA (mtDNA) and an increase in the incidence of 4977 deletion of mtDNA (mtDNA(4977)) in the UL tissue of POP patients. CONCLUSIONS: The alternations of mtDNA may play an important role in the molecular pathogenesis and process of POP formation.

Diminished vaginal HOXA13 expression in women with pelvic organ prolapse.

OBJECTIVE: Homeobox genes are transcriptional regulators that orchestrate embryonic development. The HOXA13 gene is responsible for the development of the vagina and regulates extracellular matrix constituents. We hypothesized that vaginal expression of HOXA13 may be decreased in women with pelvic organ prolapse (POP) and sought to determine if hypoestrogenism affects its expression. METHODS: Biopsy specimens were obtained from the anterior apex of the vagina from women with and without POP. Immunohistochemistry and real-time polymerase chain reaction were used to determine HOXA13 expression in premenopausal controls, in premenopausal women receiving leuprolide acetate, and in premenopausal and postmenopausal women with POP. RESULTS: HOXA13 was expressed in all specimens. HOXA13 expression was 14-fold lower in premenopausal women with prolapse than in premenopausal controls (P < 0.001). In both POP groups, HOXA13 expression was lower than in the leuprolide group (P

Changes in expression of fibulin-5 and lysyl oxidase-like 1 associated with pelvic organ prolapse.

OBJECTIVES: Pelvic organ prolapse is associated with defects in connective tissue including elastic fibers. The purpose of this study was to investigate expression of fibulin-5 and lysyl oxidase-like 1, which play an essential role in synthesis and assembly of elastic fibers in the uterosacral ligament, in samples taken from women with advanced pelvic organ prolapse compared with controls. STUDY DESIGN: Specimens were obtained prospectively during abdominal hysterectomy from 30 women with advanced pelvic organ prolapse and 30 controls matched to the study group for age and parity among postmenopausal women with benign gynecologic pathology. Expression levels of protein and mRNA of fibulin-5 and lysyl oxidase-like 1 in uterosacral ligaments were measured by Western blot analysis and real-time quantitative polymerase chain reactions. For statistical analyses, Student's t-test, chi-square test, Mann-Whitney U test, Fisher's exact test and Spearman's correlation were used. RESULTS: Significant decrease in the expression of mRNA of fibulin-5 was found in patients with prolapse (P-value=0.042; 0.743+/-0.229 and 1.061+/-0.537). The expression of mRNA of lysyl oxidase-like 1 was increased in the patient group (P-value=0.017; 4.099+/-2.832 and 1.816+/-1.602). Similar results were shown in Western blot analysis. Differences in expression of fibulin-5 and lysyl oxidase-like 1 were noted according to stage of prolapse (P-value=0.037 and 0.009). CONCLUSIONS: There was decreased expression of fibulin-5 and increased expression of lysyl oxidase-like 1 in uterosacral ligaments in patients with pelvic organ prolapse, which suggests the possibility of defects in elastin synthesis.