The Impact of Vaginal Probiotics on Pessary Use: A Randomized Controlled Trial.

IMPORTANCE: Pessary-related adverse effects are common, and treatment options are limited. Probiotics may improve pessary-related adverse effects by altering the vaginal microenvironment. OBJECTIVE: This study aimed to evaluate the effect of a vaginal probiotic suppository on the vaginal microenvironment among pessary users. STUDY DESIGN: Women who used pessaries were randomized to vaginal probiotic suppository use versus without use. The intervention was a vaginal probiotic suppository and moisturizing vaginal gel. The vaginal microenvironment was assessed using Gram stain and Nugent's criteria at baseline and 3 months by a microbiologist blinded to group allocation. Symptoms and experience with use of the probiotic were assessed using questionnaires. The primary outcome was change in lactobacilli count on Nugent subscore at 3 months. RESULTS: A total of 147 postmenopausal women were randomized (86 to the intervention arm and 61 to the control arm), and 124 (87.9%) presented for a 3-month follow-up. There was no difference between the arms in age, race, body mass index, and Charlson Comorbidity Index. A majority of participants had the pessary managed by the health care professional (intervention arm vs control arm, 46 [76.7%] vs 55 [68.8%]; P = 0.30). Composition of the vaginal microenvironment did not differ with or without probiotic treatment at 3 months. Bother from vaginal symptoms, including discharge, itching, and discomfort, did not differ between arms. Adverse effects from the intervention were minor, resolved with discontinuation, and occurred at 39.1%. CONCLUSION: Vaginal probiotic suppository use did not affect the composition of the vaginal microenvironment, patient satisfaction, or vaginal symptoms after 3 months of use in pessary users.

Risk factors for rotator cuff disease: A systematic review and meta-analysis of diabetes, hypertension, and hyperlipidemia.

BACKGROUND: Rotator cuff disease is a common cause of shoulder pain. Comorbidities such as diabetes, hypertension, and hyperlipidemia may be associated with rotator cuff disease, likely because of mechanisms related to vascular insufficiency. OBJECTIVES: We performed a systematic review of the association of diabetes, hypertension, and hyperlipidemia with the diagnosis of rotator cuff disease. METHODS: Following systematic queries of PubMed, Embase, Cochrane, CINAHL, and Science Direct, articles meeting eligibility criteria and reporting on the association of one or more risk factors (diabetes, hypertension, and hyperlipidemia) and rotator cuff disease were considered. Meta-analysis was performed to quantitatively summarize the associations between each risk factor and rotator cuff disease. We assessed study quality with the Newcastle-Ottawa Scale (NOS) and performed a qualitative assessment of risk of bias. RESULTS: After a full-text review of 212 articles, 12 articles assessing diabetes, 5 assessing hypertension and 8 assessing hyperlipidemia were eligible. The odds of having rotator cuff disease was increased with diabetes (odds ratio [OR] 1.49, 95% confidence interval [CI] 1.43-1.55), hypertension (OR 1.40, 95% CI 1.19-1.65) and hyperlipidemia/dyslipidemia (OR 1.48, 95% CI 1.42-1.55). Diabetes was also specifically associated with rotator cuff tears (OR 1.28, 95% CI 1.07-1.52). Synthesizing assessment for risk of bias suggested that current epidemiologic evidence for an association was plausible for diabetes and hyperlipidemia but not hypertension. CONCLUSIONS: Diabetes, hypertension, and hyperlipidemia were associated with rotator cuff disease in our meta-analysis. However, the possibility of bias exists for all 3 co-morbidities evaluated and is likely highest for hypertension. High-quality studies with the ability to incorporate time since first diagnosis of co-morbidity are scarce and much needed.

The Relationship of Aging and Smoking With Rotator Cuff Disease: A Systematic Review and Meta-analysis.

OBJECTIVE: Despite rotator cuff disease being one of the most common causes of shoulder pain, its pathogenesis and biology are poorly understood. In this study, we synthesized evidence from studies reporting associations for aging and smoking status in relation to rotator cuff disease. DESIGN: A systematic review was performed using multiple databases (PubMed, Embase, Cochrane, CINAHL, and Science Direct). Articles that met our eligibility criteria and presented data on the association between aging and/or smoking status and rotator cuff disease were included. We performed meta-analyses and reported cumulative effects using odds ratios and corresponding 95% confidence intervals. RESULTS: Of the 212 articles eligible for full-text review, seven studies reported on the relationship between aging and rotator cuff disease, and 10 studies reported on the relationship between smoking and rotator cuff disease. Aging was consistently associated with increased odds of having rotator cuff disease when assessed continuously (per 10-yr increase: odds ratio = 1.20, 95% confidence interval = 1.18-1.21) or categorically (ages <40 yrs vs: [a] 40-44 yrs [odds ratio = 2.71, 95% confidence interval = 1.78-4.13], [b] 45-49 yrs [odds ratio = 4.33, 95% confidence interval = 2.88-6.55], and [c] ≥50 yrs [odds ratio = 6.97, 95% confidence interval = 4.85-10.01]). Assessing studies that reported smoking status as current smokers versus nonsmokers, current smokers were more likely to have rotator cuff disease (odds ratio = 1.94, 95% confidence interval = 1.52-2.48). However, a statistically significant association was not found when never smokers were compared with former smokers (odds ratio = 1.08, 95% confidence interval = 0.97-1.20) and to current smokers (odds ratio = 0.97, 95% confidence interval = 0.87-1.07). CONCLUSIONS: In this systematic review and meta-analysis, increasing age was a strong risk factor for rotator cuff disease. The finding that current smokers are more likely to have rotator cuff disease as compared with nonsmokers implies that cessation of smoking can potentially lead to mitigation of this risk factor.

Biopsy bacterial signature can predict patient tissue malignancy.

Considerable recent research has indicated the presence of bacteria in a variety of human tumours and matched normal tissue. Rather than focusing on further identification of bacteria within tumour samples, we reversed the hypothesis to query if establishing the bacterial profile of a tissue biopsy could reveal its histology / malignancy status. The aim of the present study was therefore to differentiate between malignant and non-malignant fresh breast biopsy specimens, collected specifically for this purpose, based on bacterial sequence data alone. Fresh tissue biopsies were obtained from breast cancer patients and subjected to 16S rRNA gene sequencing. Progressive microbiological and bioinformatic contamination control practices were imparted at all points of specimen handling and bioinformatic manipulation. Differences in breast tumour and matched normal tissues were probed using a variety of statistical and machine-learning-based strategies. Breast tumour and matched normal tissue microbiome profiles proved sufficiently different to indicate that a classification strategy using bacterial biomarkers could be effective. Leave-one-out cross-validation of the predictive model confirmed the ability to identify malignant breast tissue from its bacterial signature with 84.78% accuracy, with a corresponding area under the receiver operating characteristic curve of 0.888. This study provides proof-of-concept data, from fit-for-purpose study material, on the potential to use the bacterial signature of tissue biopsies to identify their malignancy status.

Vascular leiomyosarcoma of the femoral vein: an unusual cause of a femoral triangle mass.

Lumps in the femoral triangle are common. The differential diagnosis includes lymphadenopathy and femoral hernias. Rarer causes of lumps are saphena varix, femoral artery aneurysm or a psoas abscess. We present a leiomyosarcoma of the femoral vein that presented as a femoral triangle mass.

Vaginal pH measured in vivo: lactobacilli determine pH and lactic acid concentration.

BACKGROUND: Lactic acid (protonated lactate) has broad antimicrobial activity. Vaginal lactobacilli produce lactic acid, and are known to confer protection against reproductive tract infections when they are predominant in the vaginal microbiota. Using novel ex vivo methods, we showed that cervicovaginal fluid (CVF) from women with a predominantly lactobacilli-morphotype microbiota contains significantly more lactic acid than previously thought, sufficient to inactivate reproductive tract pathogens. Here, we measured vaginal pH in vivo in 20 women with a predominantly lactobacilli-morphotype (low Nugent score) microbiota. We also investigated the in vitro production of protons (as hydrogen ions) and lactate by vaginal lactobacilli. RESULTS: The average vaginal pH in these women was 3.80 ± 0.20, and the average lactate concentration was 0.79% ± 0.22% w/v, with pH and lactate concentration tightly correlated for each sample. In vitro, lactobacilli cultured from these CVF samples reached an average pH of 3.92 ± 0.22, but the average lactate concentration was only 0.14% ± 0.06% w/v, approximately five-fold less than in the corresponding CVF samples. When the pH of the cultures was raised, lactate and hydrogen ion production resumed, indicating that production of lactate and hydrogen ions by vaginal lactobacilli is limited primarily by their sensitivity to hydrogen ion concentration (low pH) not lactate concentration. CONCLUSIONS: Some vaginal lactobacilli cultures have a lower limiting pH than others, and limiting pHs in vitro showed good correlation with pHs measured in vivo. The limiting pH of the lactobacilli predominant in a woman's vaginal microbiota seems critical in determining the concentration of antimicrobial lactic acid protecting her.

The implausible "in vivo" role of hydrogen peroxide as an antimicrobial factor produced by vaginal microbiota.

In the cervicovaginal environment, the production of hydrogen peroxide (H2O2) by vaginal Lactobacillus spp. is often mentioned as a critical factor to the in vivo vaginal microbiota antimicrobial properties. We present several lines of evidence that support the implausibility of H2O2 as an "in vivo" contributor to the cervicovaginal milieu antimicrobial properties. An alternative explanation is proposed, supported by previous reports ascribing protective and antimicrobial properties to other factors produced by Lactobacillus spp. capable of generating H2O2. Under this proposal, lactic acid rather than H2O2 plays an important role in the antimicrobial properties of protective vaginal Lactobacillus spp. We hope this commentary will help future research focus on more plausible mechanisms by which vaginal Lactobacillus spp. exert their antimicrobial and beneficial properties, and which have in vivo and translational relevance.

A simple technique for intubating the mouth during OGD in patients with previous neck radiation.

Trismus and microstomia are commonly associated complications of neck irradiation. In recent years we are seeing an increase in the number of patients with various head and neck cancers being treated with radiotherapy. This can pose a significant challenge in performing oesophagogastroduodenoscopy (OGD) in this cohort of patients. We describe a novel technique for intubating the mouth during OGD in patients with previous neck radiation. Instead of placing a standard mouthpiece, we place the barrel of a 5 mm syringe, which is cut in half, into the patient's mouth. This method allows easy passage of the gastroscope, where the mouth opening is limited by trismus from prior radiation. It also serves to protect the patient's teeth during OGD. Successful intubation with a gastroscope is possible in patients with severe trismus using our novel technique.

Chylous ascites as a consequence of idiopathic pancreatitis.

Chylous ascites (chyloperitoneum) is a rare clinical condition, characterized by an accumulation of lymph fluid in the peritoneal cavity. Most commonly it is associated with abdominal malignancy (usually lymphoma). We present an unusual case of a woman who developed a persistent pseudocyst and recurrent chylous ascites following acute necrotizing pancreatitis.

Vaginal pH and microbicidal lactic acid when lactobacilli dominate the microbiota.

Lactic acid at sufficiently acidic pH is a potent microbicide, and lactic acid produced by vaginal lactobacilli may help protect against reproductive tract infections. However, previous observations likely underestimated healthy vaginal acidity and total lactate concentration since they failed to exclude women without a lactobacillus-dominated vaginal microbiota, and also did not account for the high carbon dioxide, low oxygen environment of the vagina. Fifty-six women with low (0-3) Nugent scores (indicating a lactobacillus-dominated vaginal microbiota) and no symptoms of reproductive tract disease or infection, provided a total of 64 cervicovaginal fluid samples using a collection method that avoided the need for sample dilution and rigorously minimized aerobic exposure. The pH of samples was measured by microelectrode immediately after collection and under a physiological vaginal concentration of CO2. Commercial enzymatic assays of total lactate and total acetate concentrations were validated for use in CVF, and compared to the more usual HPLC method. The average pH of the CVF samples was 3.5 ± 0.3 (mean ± SD), range 2.8-4.2, and the average total lactate was 1.0% ± 0.2% w/v; this is a five-fold higher average hydrogen ion concentration (lower pH) and a fivefold higher total lactate concentration than in the prior literature. The microbicidal form of lactic acid (protonated lactic acid) was therefore eleven-fold more concentrated, and a markedly more potent microbicide, than indicated by prior research. This suggests that when lactobacilli dominate the vaginal microbiota, women have significantly more lactic acid-mediated protection against infections than currently believed. Our results invite further evaluations of the prophylactic and therapeutic actions of vaginal lactic acid, whether provided in situ by endogenous lactobacilli, by probiotic lactobacilli, or by products that reinforce vaginal lactic acid.

Fatal cerebral air embolism following endoscopic evaluation of rectal stump.

A 63-year-old man underwent endoscopic evaluation of the rectal stump for rectal bleeding and suffered a massive cerebral air embolism with severe neurological impairment and subsequent death. The patient underwent a Hartmann's procedure 9 month previously for ischaemic bowel and was noted to have portal hypertension at laparotomy. We hypothesise that air entered the venous plexus around rectum and entered the azygos vein via a porto-systemic shunt and travelled retrogradely via the superior vena cava to the venous sinuses of the brain.

Adenovirus-mediated transcriptional targeting of colorectal cancer and effects on treatment-resistant hypoxic cells.

BACKGROUND: Colorectal cancer is the second leading cause of cancer-related mortality and frequently presents with locally advanced or metastatic disease. Adenovirus (Ad) vectors are important gene delivery agents because they offer efficient and broad tissue transduceability. However, their ability to penetrate through multicell layers in colorectal cancers and maintain expression in colon tumor-related hypoxic conditions has yet to be analyzed. Furthermore, their broad tissue tropism presents safety concerns. MATERIALS AND METHODS: An ex vivo cultured patient tumor sample model was employed to examine Ad transduction of colorectal tumors. RESULTS: Results obtained from Ad delivery of the firefly luciferase (FLuc) reporter gene indicated that colon tumor tissue was more amenable to Ad transduction than other tumor histologic types examined (breast and ovary). Ad transduction levels were significantly higher than a range of viral and nonviral methods examined in patient colon tissue. Control of transgene expression using the CXC chemokine receptor 4 (CXCR4) promoter was examined as a strategy to confine expression to tumor cells. An Ad construct carrying FLuc under the control of the human CXCR4 promoter demonstrated low reporter gene expression compared with the ubiquitously expressing cytomegalovirus promoter in normal colon and liver tissue while providing high expression in tumors, demonstrating a 'tumour-on' and 'normal-off' phenotype in patient tissue. The effects of changing hypoxia on Ad-related transgene expression were examined in an in vitro model of hypoxic conditions relevant to clinical colorectal tumors. Reporter gene expression varied depending on the level of hypoxia, with significantly reduced levels observed with prolonged hypoxia. However, transgene expression was robust in the cycling hypoxic conditions relevant to colorectal tumors. CONCLUSION: This study provides novel, clinically relevant data demonstrating the potential for efficient gene delivery to colorectal tumors using Ad.

Evaluation of microbicide gel adherence monitoring methods.

BACKGROUND: An objective and accurate method that measures adherence to vaginal microbicide gel regimens during clinical trials could provide more accurate estimates of microbicide efficacy, aid in targeting adherence promotion resources, and enable objective assessment of adherence promotion strategies. METHODS: We evaluated 4 methods to assess whether or not gel applicators had been vaginally inserted. At the study site, 50 women inserted hydroxyethylcellulose universal placebo gel through a polypropylene vaginal applicator and handled, but did not insert a second "sham-inserted" applicator. Applicators were discarded into a container capped with a medical event monitor system (MEMS) that recorded the time and date of opening. Fifteen additional participants did likewise at 2 study site visits, and administered gel on 6 intervening days at home. Applicators were scored as inserted, or not, by direct inspection under ambient light, ultraviolet (UV) light, staining with Alcian blue, and microscopic detection of vaginal cells stained with iodine. RESULTS: Mean sensitivity/specificity of 2 readings each by 3 test readers for UV, Alcian blue, ambient light, and iodine methods were 84/83, 79/83, 76/63, and 65/80%, respectively. Sensitivity of all methods was significantly higher in applicators inserted after one or more prior insertions of gel, with the highest sensitivity (95%) obtained with UV. MEMS caps accurately recorded applicator disposal time. CONCLUSIONS: The modest accuracy of all 4 methods for applicator insertions without prior gel applications may limit their accuracy in monitoring coital regimens. However, for daily dosing regimens, MEMS monitoring and UV inspection should provide a rapid, reliable, and quantitative assessment of adherence.

In vaginal fluid, bacteria associated with bacterial vaginosis can be suppressed with lactic acid but not hydrogen peroxide.

BACKGROUND: Hydrogen peroxide (H2O2) produced by vaginal lactobacilli is generally believed to protect against bacteria associated with bacterial vaginosis (BV), and strains of lactobacilli that can produce H2O2 are being developed as vaginal probiotics. However, evidence that led to this belief was based in part on non-physiological conditions, antioxidant-free aerobic conditions selected to maximize both production and microbicidal activity of H2O2. Here we used conditions more like those in vivo to compare the effects of physiologically plausible concentrations of H2O2 and lactic acid on a broad range of BV-associated bacteria and vaginal lactobacilli. METHODS: Anaerobic cultures of seventeen species of BV-associated bacteria and four species of vaginal lactobacilli were exposed to H2O2, lactic acid, or acetic acid at pH 7.0 and pH 4.5. After two hours, the remaining viable bacteria were enumerated by growth on agar media plates. The effect of vaginal fluid (VF) on the microbicidal activities of H2O2 and lactic acid was also measured. RESULTS: Physiological concentrations of H2O2 (< 100 µM) failed to inactivate any of the BV-associated bacteria tested, even in the presence of human myeloperoxidase (MPO) that increases the microbicidal activity of H2O2. At 10 mM, H2O2 inactivated all four species of vaginal lactobacilli but only one of seventeen species of BV-associated bacteria. Moreover, the addition of just 1% vaginal fluid (VF) blocked the microbicidal activity of 1 M H2O2. In contrast, lactic acid at physiological concentrations (55-111 mM) and pH (4.5) inactivated all the BV-associated bacteria tested, and had no detectable effect on the vaginal lactobacilli. Also, the addition of 10% VF did not block the microbicidal activity of lactic acid. CONCLUSIONS: Under optimal, anaerobic growth conditions, physiological concentrations of lactic acid inactivated BV-associated bacteria without affecting vaginal lactobacilli, whereas physiological concentrations of H2O2 produced no detectable inactivation of either BV-associated bacteria or vaginal lactobacilli. Moreover, at very high concentrations, H2O2 was more toxic to vaginal lactobacilli than to BV-associated bacteria. On the basis of these in vitro observations, we conclude that lactic acid, not H2O2, is likely to suppress BV-associated bacteria in vivo.

Targeting of breast metastases using a viral gene vector with tumour-selective transcription.

BACKGROUND: Adeno-associated virus (AAV) vectors have significant potential as gene delivery vectors for cancer gene therapy. However, broad AAV2 tissue tropism results in nonspecific gene expression. MATERIALS AND METHODS: We investigated use of the C-X-C chemokine receptor type 4 (CXCR4) promoter to restrict AAV expression to tumour cells, in subcutaneous MCF-7 xenograft mouse models of breast cancer and in patient samples, using bioluminescent imaging and flow cytometric analysis. RESULTS: Higher transgene expression levels were observed in subcutaneous MCF-7 tumours relative to normal tissue (muscle) using the CXCR4 promoter, unlike a ubiquitously expressing Cytomegalovirus promoter construct, with preferential AAVCXCR4 expression in epithelial tumour and CXCR4-positive cells. Transgene expression following intravenously administered AAVCXCR4 in a model of liver metastasis was detected specifically in livers of tumour bearing mice. Ex vivo analysis using patient samples also demonstrated higher AAVCXCR4 expression in tumour compared with normal liver tissue. CONCLUSION: This study demonstrates for the first time, the potential for systemic administration of AAV2 vector for tumour-selective gene therapy.