Dysregulation of X-ray repair cross-complementing 4 expression in the eutopic endometrium of women with endometriosis.

Recent data suggest that the DNA damage response (DDR) is altered in the eutopic endometrium (EE) of women with endometriosis and this probably ensues in response to higher DNA damage encountered by the EE in endometriosis. DDR operates in a tissue-specific manner and involves different pathways depending on the type of DNA lesions. Among these pathways, the non-homologous end joining (NHEJ) pathway plays a critical role in the repair of dsDNA breaks. The present study was undertaken to explore whether NHEJ is affected in the EE of women with endometriosis. Toward this, we focused on the X-ray repair cross-complementing 4 (XRCC4) protein, one of the core components of the NHEJ pathway. Endometrial XRCC4 protein levels in the mid-proliferative phase were found significantly (P < 0.05) downregulated in women with endometriosis, compared to control women. Investigation of a microarray-based largest dataset in the Gene Expression Omnibus database (GSE51981) revealed a similar trend at the transcript level in the EE of women with endometriosis, compared to control women. Further in vitro studies were undertaken to explore the effects of H2O2-induced oxidative stress on DNA damage, as assessed by γ-H2AX and 8-hydroxy-2'-deoxyguanosine (8-OHdG) immunolocalization, and XRCC4 protein levels in endometrial stromal (hTERT immortalized human endometrial stromal cell line (ThESCs)) and epithelial (Ishikawa) cells. A significant decrease in XRCC4 protein levels and significantly higher localization of γ-H2AX and 8-OHdG were evident in ThESCs and Ishikawa cells experiencing oxidative stress. Overall, the study demonstrates that the endometrial XRCC4 expression is dysregulated in women with endometriosis and this could be due to higher oxidative stress in endometriosis.

Protocol for a case-control study investigating the clinical phenotypes and genetic regulation of endometriosis in Indian women: the ECGRI study.

INTRODUCTION: Endometriosis is one of the common, gynaecological disorders associated with chronic pelvic pain and subfertility affecting ~10% of reproductive age women. The clinical presentation, etiopathogenesis of endometriosis subtypes and associated risk factors are largely unknown. Genome-Wide Association (GWA) Studies (GWAS) provide strong evidence for the role of genetic risk factors contributing to endometriosis. However, no studies have investigated the association of the GWAS-identified single-nucleotide polymorphism (SNPs) with endometriosis risk in the Indian population; therefore, one-sixth of the world's population is not represented in the global genome consortiums on endometriosis. The Endometriosis Clinical and Genetic Research in India (ECGRI) study aims to broaden our understanding of the clinical phenotypes and genetic risks associated with endometriosis. METHODS AND ANALYSIS: ECGRI is a large-scale, multisite, case-control study of 2000 endometriosis cases and 2000 hospital controls to be recruited over 4 years at 15 collaborating study sites across India covering representative Indian population from east,north-east, north, central, west and southern geographical zones of India. We will use the World Endometriosis Research Foundation Endometriosis Phenome and Biobanking Harmonisation Project (WERF-EPHect) data collection instruments for capturing information on clinical, epidemiological, lifestyle, environmental and surgical factors. WERF-EPHect standard operating procedures will be followed for the collection, processing and storage of biological samples. The principal analyses will be for main outcome measures of the incidence of endometriosis, disease subtypes and disease severity determined from the clinical data. This will be followed by GWAS within and across ethnic groups. ETHICS AND DISSEMINATION: The study is approved by the Institutional Ethics Committee of Indian Council of Medical Research-National Institute for Research in Reproductive Health and all participating study sites. The study is also approved by the Health Ministry Screening Committee of the Government of India. The results from this study will be actively disseminated through discussions with endometriosis patient groups, conference presentations and published manuscripts.

Endometrial DNA damage response is modulated in endometriosis.

STUDY QUESTION: Is the DNA damage response (DDR) dysregulated in the eutopic endometrium of women with endometriosis? SUMMARY ANSWER: Endometrial expression of genes involved in DDR is modulated in women with endometriosis, compared to those without the disease. WHAT IS KNOWN ALREADY: Ectopic endometriotic lesions are reported to harbour somatic mutations, thereby hinting at dysregulation of DDR and DNA repair pathways. However, it remains inconclusive whether the eutopic endometrium also manifests dysregulated DDR in endometriosis. STUDY DESIGN, SIZE, DURATION: For this case-control study conducted between 2015 and 2019, eutopic endometrial (E) samples (EE- from women with endometriosis, CE- from women without endometriosis) were collected in either mid-proliferative (EE-MP, n = 23; CE-MP, n = 17) or mid-secretory (EE-MS, n = 17; CE-MS, n = 9) phases of the menstrual cycle. This study compares: (i) DNA damage marker localization, (ii) expression of DDR genes and (iii) expression of DNA repair genes in eutopic endometrial samples from women with and without endometriosis. PARTICIPANTS/MATERIALS, SETTING, METHODS: The study included (i) 40 women (aged 31.9 ± 0.81 years) with endometriosis and (ii) 26 control women (aged 31.4 ± 1.02 years) without endometriosis. Eutopic endometrial samples from the two groups were divided into different parts for histological analysis, immunohistochemistry, RNA extraction, protein extraction and comet assays. Eighty-four genes of relevance in the DNA damage signalling pathway were evaluated for their expression in eutopic endometrial samples, using RT2 Profiler PCR arrays. Validations of the expression of two GADD (Growth Arrest DNA Damage Inducible) proteins - GADD45A and GADD45G were carried out by immunoblotting. DNA damage was assessed by immunohistochemical localization of γ-H2AFX (a phosphorylated variant of histone H2AX) and 8-OHdG (8-hydroxy-2'-deoxyguanosine). RNA sequencing data from mid-proliferative (EE-MP, n = 4; CE-MP, n = 3) and mid-secretory phase (EE-MS and CE-MS, n = 4 each) endometrial samples were scanned to compare the expression status of all the genes implicated in human DNA repair. PCNA (Proliferating Cell Nuclear Antigen) expression was determined to assess endometrial proliferation. Residual DNA damage in primary endometrial cells was checked by comet assays. Public datasets were also scanned for the expression of DDR and DNA repair genes as our RNASeq data were limited by small sample size. All the comparisons were made between phase-matched endometrial samples from women with and without endometriosis. MAIN RESULTS AND THE ROLE OF CHANCE: Endometrial expression of DDR genes and intensity of immunolocalized γ-H2AFX were significantly (P < 0.05) higher in EE, compared to CE samples. DDR proteins, especially those belonging to the GADD family, were found to be differentially abundant in EE, as compared to CE. These patterns were evident in both mid-proliferative and mid-secretory phases. Intriguingly, higher DDR was associated with increased cell proliferation in EE-MP, compared to CE-MP. Furthermore, among the differentially expressed transcripts (DETs) encoded by DNA repair genes, the majority showed up-regulation in EE-MP, compared to CE-MP. Interestingly, CE-MP and EE-MP had a comparable percentage (P > 0.05) of cells with residual DNA damage. However, unlike the mid-proliferative phase data, many DETs encoded by DNA repair genes were down-regulated in EE-MS, compared to CE-MS. An analysis of the phase-matched control and endometriosis samples included in the GSE51981 dataset available in the Gene Expression Omnibus database also revealed significant (P < 0.05) alterations in the expression of DDR and DNA repair genes in EE, compared to CE. LARGE-SCALE DATA: N/A. LIMITATIONS, REASONS FOR CAUTION: The study was conducted on a limited number of endometrial samples. Also, the study does not reveal the causes underlying dysregulated DDR in the eutopic endometrium of women with endometriosis. WIDER IMPLICATIONS OF THE FINDINGS: Alterations in the expression of DDR and DNA repair genes indirectly suggest that eutopic endometrium, as compared to its healthy counterpart, encounters DNA damage-inducing stimuli, either of higher strength or for longer duration in endometriosis. It will be worthwhile to identify the nature of such stimuli and also explore the role of higher genomic insults and dysregulated DDR/DNA repair in the origin and/or progression of endometriosis. STUDY FUNDING/COMPETING INTEREST(S): The study was supported by the Department of Biotechnology and Indian Council of Medical Research, Government of India. No conflict of interest is declared.

Co-infection of malaria and early clearance of SARS-CoV-2 in healthcare workers.

Healthcare workers (HCWs) are at higher risk of contracting severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. Information regarding co-infection of SARS-CoV-2 with vector-borne diseases (malaria and dengue) is crucial especially for the countries wherein malaria and dengue are endemic. The objective was to study the prevalence, demographic, clinical presentations among HCWs with coronavirus disease 2019 (COVID-19) and to compare the viral clearance in HCWs with COVID-19 and co-infection of malaria and dengue. This retrospective study was conducted at a dedicated COVID-19 hospital, BYL Nair Charitable Hospital (NH), Mumbai, India April 6th-October 31st 2020. The SARS-CoV-2 infection in HCWs was confirmed by reverse transcription-plymerase chain reaction. Out of 491 HCWs infected with SARS-CoV-2, analysis of viral clearance was carried out in 467 HCWs over seven month periods, The prevalence of SARS-CoV-2 infection in HCWs was 13% (491 out of 3711). Out of the HCWs with COVID-19, prevalence of SARS-CoV-2 infection was higher among security guards (25%) with 1% mortality. The co-infection of malaria or dengue was reported in 31 HCWs (6.3%). The mean duration of virus clearance was longer (12 days) in symptomatic HCWs as compared to asymptomatic (8 days, p < .005). The recovery of SARS-CoV-2 infection in HCWs was faster (mean 8 days) with co-infection of malaria than without malaria (p < .005). We recommend universal testing of HCWs, to optimize staffing levels during the current pandemic as HCWs are the most precious resource. There is a need to effectively implement standard protocols for prevention of vector-borne diseases, especially in the hospital settings.

Novel Anastomotic Technique for Uterine Transplant Using Utero-ovarian Veins for Venous Drainage and Internal Iliac Arteries for Perfusion in Two Laparoscopically Harvested Uteri.

STUDY OBJECTIVE: To evaluate 2 cases of uterine transplant surgery that used utero-ovarian veins as outflow channels, internal iliac arteries for perfusion, and the organ harvest surgery performed laparoscopically. DESIGN: Case study (Canadian Task Force Classification III). SETTING: An urban, private, tertiary care hospital. PATIENTS: Two patients, ages 30 and 24years, diagnosed with absolute uterine factor infertility secondary to Mayer-Rokitansky-Küster-Hauser syndrome underwent related living donor uterine transplants; donors were their mothers with normal menses. INTERVENTIONS: Retrieval of organs through minilaparotomy and laparoscopic harvest of donor internal iliac arteries and ovarian veins. MEASUREMENTS AND MAIN RESULTS: Anastomosis was completed with bilateral donor internal iliac arteries to recipient internal iliac arteries in an end-to-end manner and with bilateral donor ovarian veins to recipient external iliac veins in an end-to-side manner. The lengths of utero-ovarian veins of both donors were 11 and 11cm on both sides; the lengths of the internal iliac arteries of both donors were 10 and 7.5cm on the left side and 10 and 6cm on the right side. The operative times for harvest surgery, bench surgery and transplant surgery were 2:40 and 3:20 hours, 34:32 and 33:30 min and 4:00 and 4:30 hours respectively for recipients 1 and 2. Daily postoperative uterine Doppler was completed through day 8 and then every other day and showed good intrauterine blood flow (i.e., low resistance arcuate vessel flow; resistance index < .5). Cervical biopsies on postoperative days 7 and 14 showed no evidence of rejection in either recipient. Both recipients started menstruating within 2 months of surgery. CONCLUSION: By using ovarian veins as outflow channels, the challenges involved in dissection along the internal iliac vein are avoided, and harvesting the donor internal iliac artery reduces the tension on vascular anastomosis. The selection of vessels to be harvested could make the technique reproducible, although larger studies are warranted to confirm results.

Laparoscopic-Assisted Uterus Retrieval From Live Organ Donors for Uterine Transplant: Our Experience of Two Patients.

STUDY OBJECTIVE: To report the first ever laparoscopic-assisted live donor uterus retrieval in 2 patients for uterus transplant. DESIGN: Case study (Canadian Task Force classification III). SETTING: Galaxy CARE Laparoscopy Institute, Pune, India. PATIENTS: Two patients with absolute uterine factor infertility with their mothers as donors. INTERVENTIONS: In vitro fertilization and uterine transplant. MEASUREMENTS AND MAIN RESULTS: A 12-member team was formed, and approval for transplant was obtained from the institutional review board. Pretransplant, in vitro fertilization for both patients was done. Two consecutive uterine transplants were done on 2 successive days. Vessels were harvested laparoscopically in both donors. Uterus and harvested vessels were retrieved by a small abdominal incision to prevent injury and infection. The uterus was transplanted in the recipients by end to side anastomosis of the harvested vessels to external iliac vessels, followed by anchoring of supports of the donor uterus to those of the recipients. Surgical intra- and postoperative parameters, postoperative investigations, and follow-up data of 6 months were measured. Operative time for laparoscopic donor surgery was 4 hours. Bench surgery took 45 minutes. Recipient surgery time was 4 hours. There were no intraoperative or immediate postoperative complications. Both the recipients started menstruating after 34 days and 48 days, respectively, and have had 6 cycles of menses at regular intervals. Uterine artery Doppler showed good flow in both patients. Hysteroscopy-guided cervical biopsies were used as a method of surveillance of graft rejection after uterine transplant. Office hysteroscopy was done after 2 months in both patients, and hysteroscopy-guided endometrial and cervical biopsies were taken. Minimal slough was seen on the endometrium in the patient with Mayer-Rokitansky-Küster-Hauser syndrome, which was removed. Repeat hysteroscopy after 10 days showed a healthy endometrium. CONCLUSIONS: Laparoscopic-assisted uterus donor retrieval is feasible and affords all the advantages of a minimally invasive technique, thereby reducing the morbidity of the procedure. It helps in better dissection of the vessels, shortens the operative time, and helps to minimize tissue handling of the harvested uterus and vessels.

Laparoscopic-Assisted Uterus Retrieval From Live Organ Donors for Uterine Transplant.

STUDY OBJECTIVE: The authors present the first ever laparoscopic-assisted uterus retrieval in a live donor for uterus transplant. DESIGN: A step-by-step surgical demonstration. SETTING: Galaxy CARE Laparoscopy Institute, Pune, India. PATIENTS: Two patients, ages 21 and 26 years, with Mayer-Rokitansky-Küster-Hauser syndrome and Asherman syndrome, respectively, with their mothers as donors. INTERVENTIONS: A 12-member team was formed. After a review of the available literature on uterine transplant, a protocol was formulated and submitted to the Institutional Review Board (IRB). Approval from the Institutional Review Board was obtained. Thorough screening of the candidates was done. Two consecutive uterine transplants were done on 2 successive days. Vessels were harvested laparoscopically in both donors. Uterus was retrieved through a small abdominal incision, to prevent any injury to the uterus and harvested vessels. Uterus was transplanted in the recipients by end-to-side anastomosis of the harvested vessels to the external iliac vessels, followed by anchoring of supports of the donor uterus to those of the recipients. MEASUREMENTS AND MAIN RESULTS: Surgical intra- and postoperative parameters, postoperative investigations, and follow-up data of 4 months. The operative time for laparoscopic donor surgery was 4 hours. Bench surgery took 45 minutes. The recipient surgery was completed in 4 hours. There were no intraoperative or immediate postoperative complications. Both recipients started menstruating after 34 days and 48 days, respectively, and have had 3 cycles of menses at regular intervals to date. After discharge, follow-up cervical biopsies at 3 weekly intervals showed no signs of rejection. Uterine artery Doppler ultrasound showed good flow in both patients. CONCLUSION: Laparoscopic-assisted donor retrieval is feasible and affords all advantages of a minimally invasive technique. It helps in better dissection of vessels, shortens the operative time, and helps minimize tissue handling, thereby reducing the morbidity of the procedure.

Panel of Autoimmune Markers for Noninvasive Diagnosis of Minimal-Mild Endometriosis.

Endometriosis, characterized by the presence of endometrial-like tissue at extrauterine sites, is a common, chronic, estrogen-dependent, inflammatory condition associated with pelvic pain, subfertility, dysmenorrhea, and dyspareunia, affecting about 10% of reproductive-age women in any population. The diagnosis of endometriosis is usually delayed on an average by 8 to 11 years leading to significant consequences in terms of disease progression. The current study was aimed to validate enzyme-linked immunosorbent assay based on the epitopes of stomatin-like protein 2, tropomodulin 3 (TMOD3), and tropomyosin 3 (TPM3) for diagnosis of minimal-mild endometriosis (revised American Fertility Society Classification (rAFS) stage I-II) and to compare the performance with the reported markers: cancer antigen (CA) 125, CA19-9, α-enolase, Serine/threonine-protein kinase (PDIK1L), and syntaxin 5. This was a cross-sectional, multicenter study conducted during the year 2012 to 2015. Women with minimal-mild endometriosis (rAFS stage I-II [n = 133]) and healthy controls (n = 104) were screened for 11 novel autoimmune markers and reported markers α-enolase, PDIK1L, syntaxin 5, CA-125, and CA19-9. The sensitivity and diagnostic accuracy of serum antibodies against all the 11 epitopes were higher than that of CA-125, CA19-9, α-enolase, PDIK1L, and syntaxin 5 for diagnosis of rAFS stage I to II endometriosis. The sensitivity of 6 biomarkers (anti-TMOD3b-autoAb, anti-TMOD3c-autoAb, anti-TMOD3d-autoAb, anti-TPM3a-autoAb, anti-TPM3c-autoAb, and anti-TPM3d-autoAb) was higher at the specificity of ≥80% for diagnosis of rAFS stage I to II endometriosis as well as ultrasound-negative endometriosis. Further, logistic regression models of this panel of biomarkers showed increase in sensitivity, specificity, and diagnostic accuracy than individual biomarkers. The panel of 6 autoimmune biomarkers could be useful in setting up of noninvasive diagnostic test for detection of minimal-mild endometriosis.

Endometriotic mesenchymal stem cells exhibit a distinct immune phenotype.

Endometriosis is a significant debilitating gynecological problem affecting women of the reproductive age group and post-menopause. Recent reports suggest a role for endometriotic mesenchymal stem cells (ectopic MSCs) in the pathogenesis of endometriosis. To investigate the plausible mechanisms leading to the pathogenic behavior of ectopic MSCs, we compared the immunomodulatory properties of eutopic (healthy) and ectopic MSCs. We analyzed MSC phenotypes, differentiation potential, differential gene expression for an array of pattern recognition receptors (PRRs) and pro-inflammatory cytokine release along with markers of migration and angiogenesis among eutopic and ectopic MSCs. Further, alterations in immunosuppressive functions of eutopic and ectopic MSCs were examined by co-culturing them with mitogen-activated allogeneic PBMCs. Transcripts of PRRs such as all Toll-like receptors (TLR1-10), except TLR8, collectins (CL-L1, CL-P1 and CL-K1), NOD-1 and NOD-2 receptors and secreted pro-inflammatory cytokines like IL-6, IFN-γ, vascular endothelial growth factor (VEGF), epidermal growth factor and MCP-1 were significantly up-regulated in ectopic MSCs. The anti-inflammatory cytokine transforming growth factor-ß showed significant down-regulation, while IL-10 showed a significant increase in ectopic MSCs. Further, ectopic MSCs showed up-regulated expression for markers of migration and angiogenesis such as matrix metalloproteinase-2 (MMP-2), MMP-3 and MMP-9 and VEGF, respectively. We report here that proliferation of PBMCs was less inhibited upon co-culture with ectopic MSCs compared with eutopic MSCs. The findings suggest that ectopic MSCs with increased levels of TLRs, collectins, pro-inflammatory cytokines and markers of migration and angiogenesis exhibit a distinct immune phenotype compared to eutopic MSCs. This distinct phenotype may be responsible for the reduced immunosuppressive property of ectopic MSCs and may be associated with the pathogenesis of endometriosis.

Proteolytic tailoring of the heat shock protein 70 and its implications in the pathogenesis of endometriosis.

OBJECTIVE: To investigate the mechanism underlying the appearance of a 20-kd HSP70 fragment and its consequences in the ectopic endometrium of endometriosis patients. DESIGN: Experimental study. SETTING: Research institute and obstetrics and gynecology clinic. PATIENT(S): Participants with (n = 18) and without (n = 20) endometriosis. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Reverse-transcription polymerase chain reaction, protease assays, and in silico tools were used to investigate the origin of the 20-kd HSP70 fragment. Immunocolocalization studies were carried out to determine whether subtilisin/kexin isozyme 1 (SKI-1) and HSP70 are colocalized. Expression and localization of surrogate markers of inflammation, such as nuclear factor NF-κB and interleukin IL-6 were examined by immunoblotting and in situ studies. RESULT(S): HSP70 is posttranslationally processed into a 20-kd fragment by SKI-1, a protease of the subtilisin family, in ectopic endometrium (ECE). Immunocolocalization studies revealed spatial proximity of SKI-1 and HSP70 in ECE. Furthermore, ECE demonstrated nuclear localization of the transcription factor, NF-κB and high expression of its target protein, IL-6. CONCLUSION(S): This study hints at the possible mechanisms underlying the trimming of HSP70 in ECE and also at the role of proteases in the pathogenesis of endometriosis. The possible repercussions of HSP70 fragmentation include dysregulation of key regulatory proteins, resulting in the escalation of inflammatory events in endometriotic lesions.

"Spot"-ting differences between the ectopic and eutopic endometrium of endometriosis patients.

OBJECTIVE: To examine proteins aberrantly expressed in the ectopic endometrium compared with eutopic endometrium from the same patient. DESIGN: Experimental study. SETTING: Research institute and an obstetrics and gynecology clinic (National Institute for Research in Reproductive Health and Sanjeevani Diagnostic Center and Maternity Home, India). PATIENT(S): Twenty participants with (n=11) and without (n=9) endometriosis. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Protein identification by two-dimensional (2D) electrophoresis and mass spectrometry as well as validation of the identified proteins by studying protein expression via Western blot and protein localization via immunohistochemical analysis. RESULT(S): Computer-assisted image analysis detected the presence of 53 protein spots in ectopic 2D gels that were conspicuous by their absence in the 2D maps of eutopic and control endometrium, i.e., these spots were detected only in ectopic gels. Eleven spots were identified by mass spectrometry. The expression of four of these proteins-haptoglobin, Rho-GDIα, SM-22α, and Rab37-have been validated by immunohistochemical and Western blotting analysis. CONCLUSION(S): This study assumes significance, as there are no reports on the comparison of the global protein profiles of paired eutopic and ectopic endometrium. Furthermore, the study demonstrates a definitive difference in the protein repertoire of the ectopic endometrium compared with its uterine counterpart in the same patient. Such studies are relevant in deciphering the complex biology of the endometriotic lesion.

Laparoscopic myomectomy: enucleation of the myoma by morcellation while it is attached to the uterus.

STUDY OBJECTIVE: To evaluate the feasibility, blood loss, length of surgery, mean hospital stay, and complications of enucleation of a myoma by morcellation while it is still attached to the uterus and to compare the technique with the standard technique of laparoscopic myomectomy. DESIGN: Randomized study (Canadian Task Force classification II-2). SETTING: Private endoscopy center. PATIENTS: Forty-four patients with symptomatic myomas confirmed by ultrasound examination were included in the study from January 2000 through December 2001 and were randomized into two groups-A and B. The inclusion criteria were the presence of a uterus larger than 12 weeks (on bimanual examination), ultrasound confirmation of the presence of at least one myoma 7 cm or greater in size, and/or presence of three or more myomas greater than 5 cm in size. INTERVENTION: The technique of laparoscopic myomectomy by enucleation of a myoma by morcellation while it is still attached to the uterus was performed in all patients in Group A. The patients in Group B underwent laparoscopic myomectomy by the conventional technique of complete enucleation of the myoma followed by morcellation. MEASUREMENTS AND MAIN RESULTS: Forty-nine myomas were removed in group A and 35 in group B. The mean weight of the myomas removed in each patient was 600.5 +/- 369.1 g in group A (95% CI 452.83-748.17 g) and 584.2 +/- 411.1 g in group B (95% CI 404.05-764.45 g) (p = .706). The mean blood loss was 283.9 +/- 229.3 mL in group A (95% CI 192.20-375.72 mL) and 218.5 +/- 110.7 mL in group B (95% CI 169.96-267.04 mL) (p = .739), the mean hospital stay was 37.91 +/- 5.44 hours in group A (95% CI 35.74-40.10 hours) and 39.5 +/- 3.634 hours in group B (95% CI 37.91-41.09 hours) (p = .236). The mean length of surgery was significantly shorter in group A (97.7 +/- 27.06 min, 95% CI 86.88-108.54 minutes) as compared with that in group B (123 +/- 38.8 min 95% CI 106.93-140.57 minutes), (p = .013). CONCLUSION: Preliminary results suggest that laparoscopic myomectomy employing the technique of enucleation of a myoma by morcellation while it is still attached to the uterus is safe and efficient. It helps to overcome certain technical difficulties inherent in the standard technique of laparoscopic myomectomy. It may help to relax the inclusion criteria of patients with myoma for laparoscopic myomectomy based on the size of the myoma.

Laparoscopic devascularization of uterine myomata followed by enucleation of the myomas by direct morcellation.

Most intraoperative conversions of laparoscopic myomectomy to laparotomy reported in the literature occur because of intraoperative bleeding. Devascularization of a uterine myoma at the start of myomectomy would help reduce the blood supply to the uterus and hence to the myoma. Another advantage of the procedure is that the need to separate the myoma from the uterus completely before morcellation, as in conventional laparoscopic myomectomy, is obviated. The tumor can be enucleated only up to about half its circumference by standard enucleation before morcellation is begun. Traction accorded by the 15-mm traumatic serrated-edge claw forceps of the morcellator during morcellation causes progressive separation of the myoma from the uterine wall, thus completing enucleation. In two patients, myomas were devascularized at the outset of myomectomy, in one by intracorporeal suturing of uterine vessels and in the other by laparoscopic bipolar coagulation of uterine vessels.

Laparoscopic removal of large multiple myomas with cumulative weight of 2.3 kg.

Large multiple myomas with a cumulative weight of 2.3 kg were removed laparoscopically from a nulliparous 32-year-old woman. The patient's recovery was uneventful, and she has been eumenorrheic in the 2 years since surgery.

Laparoscopic excision of very large myomas.

STUDY OBJECTIVE: To evaluate the feasibility, complications, and conversion rate of laparoscopic excision of very large myomas. DESIGN: Prospective study (Canadian Task Force classification II-2). SETTING: Private endoscopy center. PATIENTS: Fifty-one women with at least one myoma larger than 9 cm. INTERVENTION: Laparoscopic myomectomy. MEASUREMENTS AND RESULTS: We removed 78 myomas laparoscopically in these 51 patients. Three patients had two myomas larger than 9 cm, three had two myomas between 5 and 9 cm (in addition to 1 > 9 cm), and one had three myomas between 5 and 9 cm (in addition to 1 > 9 cm). Mean number of myomas removed/patient was 1.53 +/- 1.17 (range 1-6); 12 women (23.5%) had multiple myomectomy. The largest myoma removed was 21 cm. Mean myoma weight was 698.47 +/- 569.13 g (range 210-3400 g). Mean operating time was 136.67 +/- 38.28 minutes (range 80-270 min). Mean blood loss was 322.16 +/- 328.2 ml (range 100-2000 ml). One patient developed a broad ligament hematoma, two developed postoperative fever, and one underwent open subtotal hysterectomy 9 hours after surgery for dilutional coagulopathy. CONCLUSION: Myomectomy by laparoscopy is a safe alternative to laparotomy for very large myomas.