Birth Mode Does Not Determine the Presence of Shared Bacterial Strains between the Maternal Vaginal Microbiome and the Infant Stool Microbiome.

Dysbiosis of the neonatal gut microbiome during early life has been suggested as the missing link that may explain higher rates of certain diseases in caesarean section-delivered infants. Many studies report delivery mode-related dysbiosis in infants due to a lack of maternal vaginal microbiome exposure, prompting interventions to correct the neonatal gut microbiome by transferring these missing microbes after caesarean delivery. The maternal vaginal microbiome is among the first microbial exposures that many infants experience, yet little is known about the extent of direct transmission of maternal vaginal microbes. As part of the Maternal Microbiome Legacy Project, we aimed to determine if maternal vaginal bacteria are vertically transmitted to infants. We employed cpn60 microbiome profiling, culture-based screening, molecular strain typing, and whole-genome sequencing to determine whether identical maternal vaginal strains were present in infant stool microbiomes. We identified identical cpn60 sequence variants in both halves of maternal-infant dyads in 204 of 585 Canadian women and their newborn infants (38.9%). The same species of Bifidobacterium and Enterococcus were cultured from maternal and corresponding infant samples in 33 and 13 of these mother-infant dyads, respectively. Pulsed-field gel electrophoresis and whole-genome sequencing determined that near-identical strains were detected in these dyads irrespective of delivery mode, indicating an alternative source in cases of caesarean delivery. Overall, we demonstrated that vertical transmission of maternal vaginal microbiota is likely limited and that transmission from other maternal body sites, such as the gut and breast milk, may compensate for the lack of maternal vaginal microbiome exposure during caesarean delivery. IMPORTANCE The importance of the gut microbiome in human health and disease is widely recognized, and there has been a growing appreciation that alterations in gut microbiome composition during a "critical window" of development may impact health in later life. Attempts to correct gut microbiome dysbiosis related to birth mode are underpinned by the assumption that the lack of exposure to maternal vaginal microbes during caesarean delivery is responsible for dysbiosis. Here, we demonstrate that there is limited transmission of the maternal vaginal microbiome to the neonatal gut, even in cases of vaginal delivery. Furthermore, the presence of identical strains shared between mothers and infants in early life, even in cases of caesarean delivery, highlights compensatory microbial exposures and sources for the neonatal stool microbiome other than the maternal vagina.

Maternal vaginal microbiome composition does not affect development of the infant gut microbiome in early life.

Birth mode has been implicated as a major factor influencing neonatal gut microbiome development, and it has been assumed that lack of exposure to the maternal vaginal microbiome is responsible for gut dysbiosis among caesarean-delivered infants. Consequently, practices to correct dysbiotic gut microbiomes, such as vaginal seeding, have arisen while the effect of the maternal vaginal microbiome on that of the infant gut remains unknown. We conducted a longitudinal, prospective cohort study of 621 Canadian pregnant women and their newborn infants and collected pre-delivery maternal vaginal swabs and infant stool samples at 10-days and 3-months of life. Using cpn60-based amplicon sequencing, we defined vaginal and stool microbiome profiles and evaluated the effect of maternal vaginal microbiome composition and various clinical variables on the development of the infant stool microbiome. Infant stool microbiomes showed significant differences in composition by delivery mode at 10-days postpartum; however, this effect could not be explained by maternal vaginal microbiome composition and was vastly reduced by 3 months. Vaginal microbiome clusters were distributed across infant stool clusters in proportion to their frequency in the overall maternal population, indicating independence of the two communities. Intrapartum antibiotic administration was identified as a confounder of infant stool microbiome differences and was associated with lower abundances of Escherichia coli, Bacteroides vulgatus, Bifidobacterium longum and Parabacteroides distasonis. Our findings demonstrate that maternal vaginal microbiome composition at delivery does not affect infant stool microbiome composition and development, suggesting that practices to amend infant stool microbiome composition focus factors other than maternal vaginal microbes.

Vaginal microbiota associated with oncogenic HPV in a cohort of HPV-vaccinated women living with HIV.

BACKGROUND: Women living with HIV (WLWH) experience higher rates of human papillomavirus (HPV) infection and cervical cancer than women without HIV. Changes in the vaginal microbiome have been implicated in HPV-related disease processes such as persistence of high-risk HPV infection but this has not been well defined in a population living with HIV. METHODS: Four hundred and 20 girls and WLWH, age ≥9, across 14 clinical sites in Canada were enrolled to receive three doses of quadrivalent HPV vaccine for assessment of vaccine immunogenicity. Blood, cervical cytology, and cervico-vaginal swabs were collected. Cervico-vaginal samples were tested for HPV DNA and underwent microbiota sequencing. RESULTS: Principal component analysis (PCA) and hierarchical clustering generated community state types (CSTs). Relationships between taxa and CSTs with HPV infection were examined using mixed-effects logistic regressions, Poisson regressions, or generalized linear mixed-effects models, as appropriate. Three hundred and fifty-six cervico-vaginal microbiota samples from 172 women were sequenced. Human papillomavirus DNA was detected in 211 (59%) samples; 110 (31%) contained oncogenic HPV. Sixty-five samples (18%) were taken concurrently with incident oncogenic HPV infection and 56 (16%) were collected from women with concurrent persistent oncogenic HPV infection. CONCLUSIONS: No significant associations between taxa, CST, or microbial diversity and HPV-related outcomes were found. However, we observed weak associations between a dysbiotic microbiome and specific species, including Gardnerella, Porphyromonas, and Prevotella species, with incident HPV infection.

Aberrant stromal tissue factor localisation and mycolactone-driven vascular dysfunction, exacerbated by IL-1ß, are linked to fibrin formation in Buruli ulcer lesions.

Buruli ulcer (BU) is a neglected tropical disease caused by subcutaneous infection with Mycobacterium ulcerans and its exotoxin mycolactone. BU displays coagulative necrosis and widespread fibrin deposition in affected skin tissues. Despite this, the role of the vasculature in BU pathogenesis remains almost completely unexplored. We hypothesise that fibrin-driven ischemia can be an 'indirect' route to mycolactone-dependent tissue necrosis by a mechanism involving vascular dysfunction. Here, we tracked >900 vessels within contiguous tissue sections from eight BU patient biopsies. Our aim was to evaluate their vascular and coagulation biomarker phenotype and explore potential links to fibrin deposition. We also integrated this with our understanding of mycolactone's mechanism of action at Sec61 and its impact on proteins involved in maintaining normal vascular function. Our findings showed that endothelial cell dysfunction is common in skin tissue adjacent to necrotic regions. There was little evidence of primary haemostasis, perhaps due to mycolactone-dependent depletion of endothelial von Willebrand factor. Instead, fibrin staining appeared to be linked to the extrinsic pathway activator, tissue factor (TF). There was significantly greater than expected fibrin staining around vessels that had TF staining within the stroma, and this correlated with the distance it extended from the vessel basement membrane. TF-induced fibrin deposition in these locations would require plasma proteins outside of vessels, therefore we investigated whether mycolactone could increase vascular permeability in vitro. This was indeed the case, and leakage was further exacerbated by IL-1ß. Mycolactone caused the loss of endothelial adherens and tight junctions by the depletion of VE-cadherin, TIE-1, TIE-2 and JAM-C; all Sec61-dependent proteins. Taken together, our findings suggest that both vascular and lymphatic vessels in BU lesions become "leaky" during infection, due to the unique action of mycolactone, allowing TF-containing structures and plasma proteins into skin tissue, ultimately leading to local coagulopathy and tissue ischemia.

Inhibition of the SEC61 translocon by mycolactone induces a protective autophagic response controlled by EIF2S1-dependent translation that does not require ULK1 activity.

The Mycobacterium ulcerans exotoxin, mycolactone, is responsible for the immunosuppression and tissue necrosis that characterizes Buruli ulcer. Mycolactone inhibits SEC61-dependent co-translational translocation of proteins into the endoplasmic reticulum and the resultant cytosolic translation triggers degradation of mislocalized proteins by the ubiquitin-proteasome system. Inhibition of SEC61 by mycolactone also activates multiple EIF2S1/eIF2α kinases in the integrated stress response (ISR). Here we show mycolactone increased canonical markers of selective macroautophagy/autophagy LC3B-II, ubiquitin and SQSTM1/p62 in diverse disease-relevant primary cells and cell lines. Increased formation of puncta positive for the early autophagy markers WIPI2, RB1CC1/FIP200 and ATG16L1 indicates increased initiation of autophagy. The mycolactone response was SEC61A1-dependent and involved a pathway that required RB1CC1 but not ULK. Deletion of Sqstm1 reduced cell survival in the presence of mycolactone, suggesting this response protects against the increased cytosolic protein burden caused by the toxin. However, reconstitution of baseline SQSTM1 expression in cells lacking all autophagy receptor proteins could not rescue viability. Translational regulation by EIF2S1 in the ISR plays a key role in the autophagic response to mycolactone. Mycolactone-dependent induction of SQSTM1 was reduced in eif2ak3-/-/perk-/- cells while the p-EIF2S1 antagonist ISRIB reversed the upregulation of SQSTM1 and reduced RB1CC1, WIPI2 and LC3B puncta formation. Increased SQSTM1 staining could be seen in Buruli ulcer patient skin biopsy samples, reinforcing genetic data that suggests autophagy is relevant to disease pathology. Since selective autophagy and the ISR are both implicated in neurodegeneration, cancer and inflammation, the pathway uncovered here may have a broad relevance to human disease.Abbreviations: ATF4: activating transcription factor 4; ATG: autophagy related; BAF: bafilomycin A1; ATG16L1: autophagy related 16 like 1; BU: Buruli ulcer; CQ: chloroquine; EIF2AK3: eukaryotic translation initiation factor 2 alpha kinase 3; CALCOCO2: calcium binding and coiled-coil domain 2; DMSO: dimethyl sulfoxide; EIF2S1: eukaryotic translation initiation factor 2 subunit alpha; ER: endoplasmic reticulum; GFP: green fluorescent protein; HDMEC: human dermal microvascular endothelial cells; HFFF: human fetal foreskin fibroblasts; ISR: integrated stress response; ISRIB: integrated stress response inhibitor; MAP1LC3B/LC3B: microtubule associated protein 1 light chain 3 beta; MEF: mouse embryonic fibroblast; Myco: mycolactone; NBR1: NBR1 autophagy cargo receptor; NFE2L2: nuclear factor, erythroid 2 like 2; OPTN: optineurin; PFA: paraformaldehyde; PtdIns3P: phosphatidylinositol-3-phosphate; RB1CC1: RB1-inducible coiled coil 1; SQSTM1: sequestosome 1; TAX1BP1: Tax1 binding protein 1; ULK: unc-51 like autophagy activating kinase; UPS: ubiquitin-proteasome system; WIPI: WD repeat domain, phosphoinositide interacting; WT: wild type.

Early Neonatal Meconium Does Not Have a Demonstrable Microbiota Determined through Use of Robust Negative Controls with cpn60-Based Microbiome Profiling.

Detection of bacterial DNA within meconium is often cited as evidence supporting in utero colonization. However, many studies fail to adequately control for contamination. We aimed to define the microbial content of meconium under properly controlled conditions. DNA was extracted from 141 meconium samples and subjected to cpn60-based microbiome profiling, with controls to assess contamination throughout. Total bacterial loads of neonatal meconium, infant stool, and controls were compared by 16S rRNA quantitative PCR (qPCR). Viable bacteria within meconium were cultured, and isolate clonality was assessed by pulsed-field gel electrophoresis (PFGE). Meconium samples did not differ significantly from controls with respect to read numbers or taxonomic composition. Twenty (14%) outliers with markedly higher read numbers were collected significantly later after birth and appeared more like transitional stool than meconium. Total bacterial loads were significantly higher in stool than in meconium, which did not differ from that of sequencing controls, and correlated well with read numbers. Cultured isolates were most frequently identified as Staphylococcus epidermidis, Enterococcus faecalis, or Escherichia coli, with PFGE indicating high intraspecies diversity. Our findings highlight the importance of robust controls in studies of low microbial biomass samples and argue against meaningful bacterial colonization in utero. Given that meconium microbiome profiles could not be distinguished from sequencing controls, and that viable bacteria within meconium appeared uncommon and largely consistent with postnatal skin colonization, there does not appear to be a meconium microbiota. IMPORTANCE Much like the recent placental microbiome controversy, studies of neonatal meconium reporting bacterial communities within the fetal and neonatal gut imply that microbial colonization begins prior to birth. However, recent work has shown that placental microbiomes almost exclusively represent contamination from lab reagents and the environment. Here, we demonstrate that prior studies of neonatal meconium are impacted by the same issue, showing that the microbial content of meconium does not differ from negative controls that have never contained any biological material. Our culture findings similarly supported this notion and largely comprised bacteria normally associated with healthy skin. Overall, our work adds to the growing body of evidence against the in utero colonization hypothesis.

Evaluation of variant calling for cpn60 barcode sequence-based microbiome profiling.

Amplification and sequencing of conserved genetic barcodes such as the cpn60 gene is a common approach to determining the taxonomic composition of microbiomes. Exact sequence variant calling has been proposed as an alternative to previously established methods for aggregation of sequence reads into operational taxonomic units (OTU). We investigated the utility of variant calling for cpn60 barcode sequences and determined the minimum sequence length required to provide species-level resolution. Sequence data from the 5´ region of the cpn60 barcode amplified from the human vaginal microbiome (n = 45), and a mock community were used to compare variant calling to de novo assembly of reads, and mapping to a reference sequence database in terms of number of OTU formed, and overall community composition. Variant calling resulted in microbiome profiles that were consistent in apparent composition to those generated with the other methods but with significant logistical advantages. Variant calling is rapid, achieves high resolution of taxa, and does not require reference sequence data. Our results further demonstrate that 150 bp from the 5´ end of the cpn60 barcode sequence is sufficient to provide species-level resolution of microbiota.

One-year results of the use of endovenous radiofrequency ablation utilising an optimised radiofrequency-induced thermotherapy protocol for the treatment of truncal superficial venous reflux.

Background In previous in vitro and ex vivo studies, we have shown increased thermal spread can be achieved with radiofrequency-induced thermotherapy when using a low power and slower, discontinuous pullback. We aimed to determine the clinical success rate of radiofrequency-induced thermotherapy using this optimised protocol for the treatment of superficial venous reflux in truncal veins. Methods Sixty-three patients were treated with radiofrequency-induced thermotherapy using the optimised protocol and were followed up after one year (mean 16.3 months). Thirty-five patients returned for audit, giving a response rate of 56%. Duplex ultrasonography was employed to check for truncal reflux and compared to initial scans. Results In the 35 patients studied, there were 48 legs, with 64 truncal veins treated by radiofrequency-induced thermotherapy (34 great saphenous, 15 small saphenous and 15 anterior accessory saphenous veins). One year post-treatment, complete closure of all previously refluxing truncal veins was demonstrated on ultrasound, giving a success rate of 100%. Conclusions Using a previously reported optimised, low power/slow pullback radiofrequency-induced thermotherapy protocol, we have shown it is possible to achieve a 100% ablation at one year. This compares favourably with results reported at one year post-procedure using the high power/fast pullback protocols that are currently recommended for this device.

Pelvic venous reflux in males with varicose veins and recurrent varicose veins.

Objectives To report on a male cohort with pelvic vein reflux and associated primary and recurrent lower limb varicose veins. Methods Full lower limb duplex ultrasonography revealed significant pelvic contribution in eight males presenting with bilateral lower limb varicose veins. Testicular and internal iliac veins were examined with either one or a combination of computed tomography, magnetic resonance venography, testicular, transabdominal or transrectal duplex ultrasonography. Subsequently, all patients received pelvic vein embolisation, prior to leg varicose vein treatment. Results Pelvic vein reflux was found in 23 of the 32 truncal pelvic veins and these were treated by pelvic vein embolisation. Four patients have since completed their leg varicose vein treatment and four are undergoing leg varicose vein treatments currently. Conclusion Pelvic vein reflux contributes towards lower limb venous insufficiency in some males with leg varicose veins. Despite the challenges, we suggest that pelvic vein reflux should probably be investigated and pelvic vein embolisation considered in such patients.

Implication of foam sclerosant inactivation by human whole blood in a laboratory setting.

Background During sclerotherapy, it has been recommended to confirm intravenous placement of the needle by aspirating blood into the sclerosant syringe. This may inactivate some, or all of the sclerosant. Aims To quantify the volume of human blood needed to completely inactivate 1 ml of sodium tetradecyl sulphate, and comparing fresh blood and blood that has been stored in an ethylenediaminetetraacetic acid tube. Methods A series of manual titrations were carried out following a procedure developed at STD Pharmaceutical Products Ltd (Hereford, UK) and listed in the British Pharmacopeia. Three percent of sodium tetradecyl sulphate stock solutions were made with increasing volumes of blood and titrated against benzethonium chloride to determine the active concentration (% w/v) of sodium tetradecyl sulphate remaining in the solution. Results A calculated approximation showed 0.3 ml of blood is required to fully inactivate 1 ml of 3% sodium tetradecyl sulphate when made into a foam. A comparison was made between the use of fresh blood and blood stored in ethylenediaminetetraacetic acid tubes. Blood stored in ethylenediaminetetraacetic acid tubes showed more inactivation of sodium tetradecyl sulphate, but this was not significant at the P ≤ 0.05 level. Conclusion The data from our study have shown that a minimum of 0.3 ml of fresh blood is required to inactivate 1 ml of 3% sodium tetradecyl sulphate as a foam and it is not significantly affected by storing blood in an ethylenediaminetetraacetic acid tube. Our methodology suggests that during foam sclerotherapy treatment, blood should not be aspirated into the syringe to confirm position, and that ultrasound guidance is more appropriate for needle placement.

Anatomical abnormalities of the pelvic venous system and their implications for endovascular management of pelvic venous reflux.

Background Pelvic venous reflux is often treated with pelvic vein embolisation; however, atypical pelvic venous anatomy may provide therapeutic challenges. Methods We retrospectively reviewed seven patient files and reported symptoms, diagnostic imaging, aberrant anatomy and means by which the interventional radiologist successfully completed the procedure. Any follow-up data were included if available. Results Four anatomical abnormalities were found: internal iliac veins draining into the contralateral common iliac vein, duplicated inferior vena cava, reverse-angle renal veins with atypical left ovarian vein drainage and direct drainage of the internal iliac vein to the inferior vena cava. All patients were successfully treated with pelvic vein embolisation. Conclusion Abnormal embryologic development may cause variable pelvic venous anatomy. Knowledge of this will enable interventional radiologists to successfully treat such patients.

Mechanochemical ablation causes endothelial and medial damage to the vein wall resulting in deeper penetration of sclerosant compared with sclerotherapy alone in extrafascial great saphenous vein using an ex vivo model.

BACKGROUND: Nonthermal, tumescentless devices are the next generation of minimally invasive devices to treat varicose veins. We aimed to investigate the effects of mechanochemical ablation (MOCA) using ClariVein (Vascular Insights, Quincy, Mass) on ex vivo great saphenous vein with histology and immunofluorescent staining. METHODS: Extrafascial great saphenous veins were harvested during surgery for varicose veins and were treated ex vivo for 10 to 11 minutes with either liquid sclerotherapy or the use of ClariVein, with and without 3% sodium tetradecyl sulfate. Veins were sectioned and subjected to hematoxylin and eosin staining and immunofluorescent staining for endothelial and smooth muscle cell markers (CD31 and α-actin) to assess overall damage and cell death in the vein wall compared with control sections. RESULTS: Histologic observations confirmed intimal damage from ClariVein, as has been previously shown; however, medial damage was also evident, which was not observed in control or liquid sclerotherapy sections. Immunofluorescent staining in the three sections studied showed a 42% decrease in CD31 staining and 27% mean reduction in α-actin staining up to a depth of 300 µm with liquid sclerotherapy. This cytotoxic effect was significantly enhanced by MOCA with a reduction in CD31 staining just above 60% and a 46% mean decrease in α-actin staining noted up to a depth of 300 µm. Far greater reductions in staining compared with sclerotherapy were observed up to a depth of 600 µm. CONCLUSIONS: MOCA using 3% sodium tetradecyl sulfate increases the penetration of the sclerosant and its effect into the vein wall and shows superior rates of tissue destruction compared with liquid sclerotherapy alone. In this model, it appears not solely to damage the endothelium but also to shear the medial layer, creating small lesions into which sclerosant can flow and exert its cytotoxic effect.

Radiofrequency-induced thermotherapy (RFiTT) in a porcine liver model and ex vivo great saphenous vein.

AIMS: To investigate the thermal spread achieved in porcine liver when using an optimised radiofrequency ablation protocol and correlate findings with the effects seen in ex vivo great saphenous vein (GSV), in order to justify clinical use with the new treatment protocol. MATERIAL AND METHODS: Porcine liver and GSV sections were treated with radiofrequency-induced thermotherapy (RFiTT) using the following settings: 20 W at 1 s/cm (linear endovenous energy density; LEED 20 J/cm), 18 W at 1 s/cm (LEED 18 J/cm), 18 W at 3 s/cm (LEED 54 J/cm), 6 W interrupted pull-back 6 s stationary every 0.5 cm (LEED 72 J/cm). Thermal spread in the liver was measured via digital imaging. GSV sections were sent to an independent laboratory for histological analysis. Previous work suggests a thermal spread of >0.65 mm in liver correlates with transmural thermoablation of a GSV. RESULTS: Parameters giving a LEED of 72 J/cm produced the best results, with a clear transmural effect in the GSV and maximal thermal spread of 1.65 mm, without excessive thermal damage or carbonisation in the ablation tract. CONCLUSIONS: Our porcine liver model correlated well with histological findings and was representative of the thermoablative effects observed in the GSV wall treated with RFiTT. Clinical investigations are now being carried out to investigate the efficacy of this protocol in the clinical setting.

The effect of a subsequent pregnancy after transjugular coil embolisation for pelvic vein reflux.

Background Pelvic venous reflux has been proven to contribute to the development of primary and recurrent varicose veins, vulval/labial varicose veins and pelvic congestion syndrome. It is associated with lower limb varicose veins in 20% of patients who have a history of at least one prior vaginal delivery. Pelvic vein embolisation is known to be a safe and effective treatment for the abolition of pelvic venous reflux. However, the effect of a subsequent pregnancy on a previously embolised patient remains largely unknown. This study aims to report the effect of pregnancy on patients that have undergone pelvic vein embolisation. Methods Patients that had previously undergone pelvic vein embolisation for pelvic venous reflux at our unit were sent a questionnaire asking if they had had a pregnancy and subsequently delivered post-embolisation. Patients responding positively were invited to attend our unit for transvaginal duplex ultrasonography of their pelvic veins. Post-pregnancy transvaginal duplex ultrasonography results were compared to pre-embolisation and 6-week post-embolisation scans. Results Eight women, aged 32-48 years (mean 38.8), were retrospectively analysed. Parity prior to embolisation ranged from 1 to 5 (mean 2.8). Initial outcomes at 6 weeks Pelvic venous reflux was completely eliminated in five patients, two patients achieved complete elimination of truncal reflux with very minor vulval reflux and one patient had persistent, mild reflux in the right internal iliac vein. Post-pregnancy outcomes Pelvic venous reflux was completely eliminated in three patients and five patients displayed pelvic venous reflux in at least one truncal vein, with or without concurrent vulval reflux. No patient showed any coil displacement or embolisation as a result of the pregnancy. Conclusions Pregnancy is associated with recurrent reflux in the pelvic veins in women who had previously been treated with coil embolisation. Following recovery from pregnancy, repeat embolisation can eliminate recurrent reflux. Pregnancy appears to be safe following coil embolisation of pelvic veins.

Pelvic congestion syndrome masquerading as osteoarthritis of the hip.

OBJECTIVES: Pelvic congestion syndrome (PCS) is associated with pelvic vein reflux (PVR), occasionally secondary to venous compression. Its symptoms, usually intra-pelvic, are alleviated following the abolition of this reflux by pelvic vein embolisation (PVE). The objective of this report is to present two cases of left hip pain, erroneously diagnosed as osteoarthritis, which disappeared after successful PVE and abolition of PVR. METHODS: Two females presented with lower limb varicose veins, and also had a history of left-sided hip pain. Both had previously been investigated for the hip pain and diagnosed as osteoarthritis despite minimal arthritic changes on pelvic X-rays. During investigation for lower limb varicose veins, both showed a pelvic origin for their leg veins and hence underwent transvaginal duplex ultrasound. This revealed PVR, and PVE was planned in both patients. RESULTS: Both patients underwent PVE and reported 'miraculous' resolution of left hip pain and also PCS symptoms including pelvic pain, irritable bowel issues and the disappearance of pelvic dragging, with almost immediate disappearance of vulval and vaginal varicosities. One patient also noted reduced clitoral sensitivity. CONCLUSION: Manifestations of PCS may vary in terms of intra- or extra-pelvic signs. PCS and PVR should be considered in the differential diagnosis of patients with arthritic symptoms in the hip without evident radiographic evidence.

Long-term results of transjugular coil embolisation for pelvic vein reflux - Results of the abolition of venous reflux at 6-8 years.

BACKGROUND: Pelvic venous reflux is known to be associated with lower limb varicose veins in 20% of women with a history of at least one previous vaginal delivery. Pelvic vein embolisation with coils has been shown to be a successful treatment in the short term. The objective of this study was to ascertain the long-term outcomes of pelvic vein embolisation for pelvic venous reflux. METHODS: Patients who had undergone pelvic vein embolisation in 2005-2007 were invited back to a specialist vein unit for transvaginal duplex ultrasonography in the summer of 2013. A total of 110 women were contacted. Pre-embolisation transvaginal duplex ultrasonography results were compared to those obtained six weeks post-procedure and at long-term follow-up. RESULTS: Twenty-eight female patients aged 40 to 75 years (mean 53.5) attended (response rate 25.5%), with parity prior to embolisation ranging from 1-5 children (mean 2.8). Mean follow-up time was 7.5 years. Six weeks post-procedure, 25 women had complete or virtual elimination of all reflux, and three had persistent reflux in at least one vein. At long-term follow-up, 11 women had complete elimination of all reflux, seven had elimination of all truncal reflux but minor reflux in vulval veins, six had minor reflux in one truncal vein, and four had significant reflux in one or more truncal veins (one of these gave birth one-year post-pelvic vein embolisation and another had coils removed during gynaecological surgery). CONCLUSIONS: Transjugular pelvic vein embolisation is a durable technique for the abolition of reflux in the pelvic veins and is particularly adept at treating reflux in the ovarian veins.

The effects of environmental and compositional manipulations on the longevity of Tessari-made foam for sclerotherapy.

BACKGROUND: Tessari-made foam sclerotherapy is performed around the world in a variety of clinics differing in methods, equipment, temperatures, and altitudes. We investigated how the following factors affected the foam's longevity: silicone vs nonsilicone syringes, volume of foam made, ratio of gas to sclerosant, use of air vs 50:50 mixture of carbon dioxide and oxygen, temperature, altitude, and 10 consecutive reuses of the syringes. METHODS: Sclerosant foam was made by the Tessari double-syringe technique. To calculate the longevity, the time was taken for half of the original volume of sclerosant to settle. Half-lives were compared with use of silicone and silicone-free syringes to make the foam. We investigated how the volume (5 mL vs 2 mL) and different ratios affected the foam by observing the half-life of 4:1, 3.5:1, and 3:1 ratios of gas to sclerosant. Air and a 50:50 mixture of carbon dioxide and oxygen were both used as the gas in changing the ratio and volume to see which produced better foam. These experiments were conducted at room (23.9°C) and refrigerator (3°C) temperatures with a constant pressure. The different ratio, volume, and silicone vs nonsilicone syringe experiments were all repeated at 9314, 7460, 4575, and 2326 feet above sea level in addition to the baseline experiment, which took place at 236 feet above sea level. To test how consecutive uses of syringes affected the foam, we made consecutive batches of foam reusing each pair of syringes 10 times; this was repeated five times with silicone syringes and twice with nonsilicone syringes. RESULTS: Switching to nonsilicone syringes can increase longevity by 70%. A larger volume of foam and a 3:1 ratio produced longer half-lives at all temperatures and altitudes. The lower (3°C) temperature increased the longevity of foam in all instances, as did the use of air. A high altitude (low pressure) had a detrimental effect on the foam's longevity. Ten consecutive syringe uses had no significant impact on the foam's half-life (silicone syringe mean between first five and last five uses, P = .95). CONCLUSIONS: The optimum conditions for making foam are nonsilicone syringes, larger volumes, a 3:1 air to sclerosant ratio, and low temperatures. Silicone syringes can be reused until friction becomes a burden. Temperature has a bigger effect than altitude on longevity of the foam. Making foam in larger volumes would allow the foam to last longer. To compensate for high altitudes (low pressures), decreasing the temperature will increase the foam's longevity.

Histologic findings after mechanochemical ablation in a caprine model with use of ClariVein.

OBJECTIVE: The use of foam and liquid sclerotherapy for the treatment of varicose veins and underlying venous reflux is widespread. A novel device, the ClariVein Occlusion Catheter (Vascular Insights LLC, Madison, Conn), has been the subject of several clinical trials in humans. We report the initial histologic results obtained with use of the device in a caprine vein model. METHODS: A total of 11 male goats (12 veins) underwent minimally invasive procedures. Unilateral mechanochemical ablation of the lateral saphenous vein by the ClariVein Occlusion Catheter with an E-140° tip was performed under fluoroscopic guidance in five veins with 5 mL of 1.5% sodium tetradecyl sulfate (STS) and in one vein with 5 mL of 0.9% saline. The remaining six received injection sclerotherapy with 5 mL of 1.5% STS or 0.9% saline. All subjects were assessed with ultrasound before the procedure and intermittently afterward during a period of 12 weeks. Subsequent termination was immediately followed by necropsy and histologic examination of the treated veins. RESULTS: Complete occlusion of the lateral saphenous vein was observed in all subjects treated with ClariVein and STS, whereas complete patency was noted in all other treatment modalities. Histologic staining with hematoxylin and eosin and Masson trichrome stain revealed total fibrotic sealing with extensive collagen production in all ClariVein/STS veins. A statistical significance was observed in the difference in the number of occluded veins between subjects treated with ClariVein/STS and those treated by injection sclerotherapy (Fisher exact test, P < .01). CONCLUSIONS: The ClariVein Occlusion Catheter with 1.5% STS can be used to achieve complete mechanochemical ablation of the lateral saphenous vein in a caprine model. The evidence in this report can be used to justify the device's use for the treatment of the great saphenous vein in subsequent human clinical trials.

Pelvic venous reflux is a major contributory cause of recurrent varicose veins in more than a quarter of women.

BACKGROUND: Leg varicose veins are associated with pelvic venous reflux in approximately 20% of women who have had children. However, most venous units do not routinely look for pelvic venous reflux or treat it. We aimed to investigate what proportion of patients with recurrent varicose veins and a history of open surgery have pelvic venous reflux as a major contributing cause of their recurrence. METHODS: A retrospective study was performed of all patients referred in the previous year with recurrent varicose veins or venous reflux disease who had previously had open surgery performed elsewhere. All patients had routine lower limb venous duplex ultrasonography, and those found to have reflux of pelvic origin underwent transvaginal duplex ultrasonography. Each case was assessed by a consultant vascular surgeon, and the major cause (or causes, if more than one) of the recurrent varicose veins was noted. RESULTS: A total of 109 patients with recurrent varicose veins in 172 legs were analyzed (mean age, 53.9 years; female-to-male ratio, 97:12). Patients were divided into four groups: group 1, all patients; group 2, female patients; group 3, female patients with children; and group 4, female patients with children who had not had hysterectomy. Pelvic venous reflux was found to be a major contributing cause of recurrent varicose veins in 44 of 172 legs (25.6%). This rose to 43 of 154 legs (27.9%) in group 2, 40 of 131 legs (30.5%) in group 3, and 37 of 111 legs (33.3%) in group 4. CONCLUSIONS: Pelvic venous reflux is a major contributing cause of recurrent varicose veins after open surgery that has rarely been reported previously. In view of this finding, we suggest that a duplex ultrasound protocol, incorporating a transvaginal duplex examination of the ovarian and internal iliac veins, be adopted for the investigation of pelvic venous reflux in female patients presenting with symptomatic leg varicose veins with duplex-observed reflux entering the leg vein pattern from the pelvis. In the event that it is found, we suggest that treatment and resolution of this source of venous reflux be considered before any intervention for the leg varicose veins, surgical or otherwise.