High serum AMH inhibits pathological growth of the low biomass endometrial microbiome.

OBJECTIVE: Although host microbiome play a role in both hormonal status and fertility, this issue has not yet been clarified. Since the endometrium is a sterile tissue, it is accepted that microbiota does not grow under normal conditions. The aim of the study was to reveal the characteristics of endometrial microbiota according to serum AMH levels in women with implantation failure. PATIENTS AND METHODS: Forty-five women aged 20-30 years with two or more implantation failures were included in the study. They were divided into 3 groups according to their serum AMH values: Group 1 -AMH <1.3 ng/ml; Group 2 - AMH between 1.3-2.6 ng/ml; Group 3 - AMH >2.6 ng/ml. Twenty-two healthy fertile women who were the same age as the infertile group and applied for cervical smear screening were accepted as the control group. Following the embryo transfer, the tip of the catheter was inserted into the transport medium under sterile conditions. Sowing was carried out by touching the tips of the catheter to the blood agar medium. After the evaluation of the petri dishes at the end of 48 hours of incubation, colonies were stained with Gram stain. Microorganisms in the colonies were identified with the Vitek-2 device according to their gram-staining characteristics and their antibiograms were made. RESULTS: A negative correlation was detected between low AMH values and the microbiome detection rates in endometrial cultures. In patients with low serum AMH levels, the chance of endometrial microbiota growth was higher in the endometrial culture medium. The most common bacteria were found to be MSSA, MRKNS and lactobacillus. Clinical pregnancy rates were found to be significantly higher in the group with high AMH levels. As AMH levels increased, positive flora detection rates decreased, while clinical pregnancy rates increased. CONCLUSIONS: Low serum AMH level increases the rate of positive endometrial microbiome in culture and decreases clinical pregnancy rates.

Serum 25-hydroxyvitamin D correlates with endometrial HOXA10 mRNA expression.

OBJECTIVE: Endocrine Society classified patients with serum 25-hydroxyvitamin D [25(OH)D] levels below 20 ng/ml as deficiency, 20-30 ng/ml as insufficiency, and >30 ng/ml as replete. This study was planned to investigate the relationship between serum vitamin D level and homeobox 10 mRNA expression in women with polycystic ovary syndrome (PCOS). PATIENTS AND METHODS: Thirty women with PCOS who failed the first IVF/ICSI attempt and were decided to have endometrial injury before second attempt were included in the study. Before the endometrial injury, the serum vitamin D levels of the women were measured, and they were divided into three equal groups as proposed by the Endocrine Society. Group 1 consisted of vitamin D deficient women (<20 ng/mL), Group 2 consisted of vitamin D insufficient women (20-30 ng/mL), and Group 3 consisted of vitamin D replete women (>30 ng/mL). Women in each group were injured with a Pipelle cannula during mid-luteal phase. Endometrial samples collected during injury were analyzed for HOXA10 mRNA expression by RT-PCR and correlated with serum vitamin D level. RESULTS: When analyzing the results according to different vitamin D thresholds, as proposed by the Endocrine Society, HOXA10 mRNA expression was comparable between vitamin D deficient and vitamin D insufficient women. The HOXA10 mRNA expression of vitamin D replete women(Group 3) was found to be higher than both vitamin D deficient (Group 1) and vitamin D insufficient women (Group 2). HOXA10 mRNA expression of the women in Group 3 was 3.3-fold higher than Group 1 and 2.6-fold higher than Group 2. HOXA10 mRNA expression was correlated to the levels of vitamin D in the Group 3 (r=0.655, p=0.02). There was no significant correlation between serum vitamin D levels and endometrial HOXA10 mRNA expression of women in both Group 1 (r=0.343, p=0.06) and Group 2 (r=0.456, p=0.08). CONCLUSIONS: Endometrium of women with PCOS with sufficient serum vitamin D levels express significantly higher HOXA10 mRNA than patients with low serum vitamin D levels.

Clomiphene citrate changes metabolite content of follicular fluid of PCOS women.

OBJECTIVE: To determine whether clomiphene citrate (CC) treatment affects the metabolite contents of a dominant follicle in polycystic ovary syndrome (PCOS). PATIENTS AND METHODS: Twenty non-obese primary infertile anovulatory PCOS women undergoing CC treatment and ten fertile women were enrolled. 6 out of 20 patients had impaired glucose tolerance test (IGT). CC was given at a dose of 150 mg on days 3-7 of cycles in the first group. 10 fertile women with a regular menstrual cycle and normal ovaries during ultrasound examination were accepted as control group. They were not given any drug for ovarian stimulation. Transvaginal sonography and follicular tracking were done to each group of participant. Both groups of subjects underwent magnetic resonance spectroscopy when the dominant follicle with a mean diameter of at least 16-18 mm was detected. Lactate (Lac), N-acetylaspartate (NAA), creatine 1 (Cr1), creatine 2 (Cr2) and choline (Cho) signal of dominant follicles were measured. Peak of each metabolite was measured in units. Voxels were placed in the center of dominant follicle. RESULTS: Compared to control group significantly decreased Cho signal was found in follicular fluid of PCOS subjects taking clomiphene. Almost three-fold decline in Cho signal was detected in PCOS group compared to Cho signal of control group (0.64±1.01 vs. 2.01 ±1.13). On the other hand, significantly increased Lac signal was detected in the dominant follicle of PCOS subjects taking clomiphene compared to control group. Almost 2 fold increase in Lac signal was noted after clomiphene treatment (1.90±0.32 vs. 0.93±2.21). The results of spectroscopy signals obtained from PCOS subjects without IGT and PCOS subjects with IGT were similar. CONCLUSIONS: Unbalanced production of Cho signal in the follicular fluid may have occurred secondary to membrane damage of cumulus-oocyte-complexes due to CC therapy.

AFC vs. AMH: prediction of ovarian response in women with endometrioma undergoing controlled ovarian stimulation.

OBJECTIVE: To assess the clinical value of antral follicle count (AFC) and anti-Mullerian hormone (AMH) for the prediction of ovarian response in women with endometrioma undergoing controlled ovarian stimulation for IVF using GnRH antagonist treatment. PATIENTS AND METHODS: Fifty patients with endometrioma who underwent their first IVF/ICSI cycle with GnRH antagonist treatment were included in the study. The average AMH values were recorded as 1.5-2 ng/mL. Fifty infertile women are not suffering from endometrioma were selected from those with male factor infertility as control. They were matched according to both serum AMH levels and age. Serum samples have been collected before the IVF treatment for determining AMH levels in both groups of subjects. Likewise, each group of subject underwent ultrasound scan for AFC on day 3. Total number of oocytes retrieved during OPU, the number of transferred embryo, implantation and clinical pregnancy rates, live birth and abortion rates, total dose of rhFSH were noted in both groups of subjects. RESULTS: Day 3 AFC was significantly higher in the control group compared to women with endometrioma. Both the number of retrieved oocytes during oocyte pick-up, MII oocytes and 2 PN embryo were significantly lower in the endometrioma. Likewise, the fertilization, implantation, clinical pregnancy and live birth rates of endometrioma group were significantly lower than those in the control group. The total rFSH dose was higher in the endometrioma group than those in control. The percentage of abortion in the endometrioma group was found to higher compared to those with controls. CONCLUSIONS: AFC is more sensitive than the AMH in detecting ovarian response in women with ovarian endometrioma. The individualization of GnRH antagonist protocols in subjects having endometrioma might be improved by using an AFC-tailored approach instead of AMH.

Disulfiram, as a candidate NF-κB and proteasome inhibitor, prevents endometriotic implant growing in a rat model of endometriosis.

OBJECTIVE: Disulfiram (DSF) exerts its therapeutic effects through oxidative, proteasome, and nuclear factor kappa beta (NF-κB) pathways. The study was planned to test the impact of DSF on growing of endometriotic implants in rats with experimentally induced endometriosis. PATIENTS AND METHODS: Thirty rats were labeled as the control (n = 8), sham (n = 6), GnRH-agonist (n = 8) and the DSF (n = 8) groups. The rats in the group 3 exposed to single dose leuprolide acetate. The rats in group 4 were treated with DSF for 21 days. The serum activity of oxidant and antioxidant markers, total oxidant status (TOS), total antioxidant status (TAS), interleukin-1ß, and tumor necrosis factor-α (TNF-α) were determined. Implants were processed for NF-κB, PCNA, and CD34 immunostaining. RESULTS: The serum concentration of malondialdehyde in the DSF group was significantly higher than those in other groups. The concentration of TAS, TNF-α, and interleukin-1ß in the DSF group considerably decreased compared to control group. Following treatment with DSF while the percentage of Grade 1 and 2 implants increased the percentage of Grade 3 and 4 implants decreased. The implants disappeared totally in two cases in the DSF group and one case in the GnRH-agonist group. The mean H-Scores of implant NF-κB and PCNA in DSF treated animals were found to significantly lower than those of the control group. CONCLUSIONS: By decreasing NF-κB expression, angiogenesis, and cell proliferation DSF prevents the growth of endometriotic implants.

Disturbed endometrial NF-κB expression in women with recurrent implantation failure.

OBJECTIVE: This study was planned to investigate whether expression levels of endometrial NF-κB1 and NFκB p65 changes in women with recurrent implantation failure (RIF). PATIENTS AND METHODS: The study group consists of 30 RIF patients having at least three previous failed IVF cycles. The control group comprises of 30 patients having one or no previous failed attempt. Endometrial samples were obtained from all participants during hysteroscopy at the late follicular phase. Samples underwent ELISA analysis and immunohistochemical staining. The semi-quantitative H-Score method was used for analyzing the intensity of endometrial NF-κB p65 expression. RESULTS: The concentrations of endometrial NF-κB1 were found to be significantly increased when compared to control subjects. Likewise, significantly increased NF-κB p65 immunoreactivity was detected in the cytoplasm of luminal and glandular epithelial cells. The H-Score of NF-κB p65 in RIF women was found to be significantly increased when compared to control group. CONCLUSIONS: Increased levels of NF-κB1 and NF-κB p65 in the endometrium of RIF women can disturb physiological inflammation which is known to be positive modulator of endometrial receptivity.

Gradient coil design considerations for iron core interventional magnets.

The requirements for access and imaging performance compete in the design of open-concept MR magnets and gradient coils. We conducted a theoretical and experimental investigation of gradient coil design using both solid and laminated pole piece construction to determine whether adequate eddy current control can be obtained without shielded gradient coils while maintaining good patient access and high gradient performance. Eddy currents, gradient characteristics, gradient efficiency, and magnet openness are compared and contrasted for various construction options based on a compact, .27 T iron yoke magnet. The resulting pole pieces and gradient coils have high efficiency for an interventional open-configuration magnet while taking up minimal space between the poles for improved patient access.

Quantification of volumetric coronary blood flow with dual-energy digital subtraction angiography.

BACKGROUND: As a solution to the well-documented problems associated with visual interpretation of coronary arteriograms, more physiological methods of assessing coronary artery stenosis are being investigated. Volumetric coronary blood flow (BF) can be a valuable aid in the analysis of functional significance of arterial obstruction. METHODS AND RESULTS: The left anterior descending coronary artery (LAD) of 15 anesthetized pigs (40 to 50 kg) was dissected free from the epicardium in its proximal portion, and a transmit-time ultrasound flow probe of the appropriate size was applied. A vascular occluder was positioned distal to the flow probe for flow adjustments. Contrast injections (2 to 4 mL/s for 3 seconds) were made into the left main coronary artery during image acquisition with a motion-immune dual-energy digital subtraction angiography (DE DSA) system. Tissue-suppressed energy-subtracted images were used to generate time-density curves. BF measurements were made in the LAD vascular bed with use of the time-density curve, with consideration that blood was momentarily replaced with contrast during the injection. In 19 comparisons, the mean BF, measured with the use of DE DSA, correlated extremely well with the mean ultrasound flow (DE DSA=0.90 ultrasound+3.10 mL/min, r=.96). Also, contrast injection increased the BF by an average of only 15% during the image-acquisition time interval. CONCLUSIONS: Accurate BF measurements can be made with motion-immune DE DSA. The BF measurements can be completed before the onset of significant changes in BF due to contrast injection. Furthermore, it is possible to make the BF measurements during routine coronary arteriography.

In-vivo validation of videodensitometric coronary cross-sectional area measurement using dual-energy digital subtraction angiography.

Previous studies indicate that conventional geometric edge detection techniques, used in quantitative coronary arteriography (QCA), have significant limitations in quantitating coronary cross-sectional area of small diameter (D) vessels (D < 1.00 mm) and lesions with complex cross-section. As a solution to this problem, we have previously reported on an in-vitro validation of a videodensitometric technique that quantitates the absolute cross-sectional area including small vessel diameter (D < 1.00 mm) and any complex shape of the vessel cross-section. For in-vivo validation, plastic tubing (5-8 mm long) with different shape complex cross-section with known cross-sectional area (A = 0.8-4.5 mm2) were percutaneously wedged in the coronary arteries of anesthetized pigs (40-50 kg). Contrast material injections (6-10 ml at 2-4 ml/sec) were made into the left main coronary artery during image acquisition using a motion immune dual-energy subtraction technique, where low and high X-ray energy and filtration were switched at 30 Hz. A comparison was made between the actual and measured cross-sectional area using the videodensitometry and edge detection techniques in tissue suppressed energy subtracted images. In eighteen comparisons the videodensitometry technique produced significantly improved results (slope = 0.87, intercept = 0.24 mm2, r = 0.94) when compared to the edge detection technique (slope = 0.42, intercept = 1.99 mm2, r = 0.39). Also, a cylindrical vessel phantom (D = 1.00-4.75 mm) was used to test the ability to calculate and correct for the effect of the out of plane angle of the arterial segment on the cross-sectional area estimation of the videodensitometry technique. After corrections were made for the out of plane angle using two different projections, there was a good correlation between the actual and the measured cross-sectional area using the videodensitometry technique (slope = 0.91, intercept = 0.11 mm2, r = 0.99). These data suggest that it is possible to quantitate absolute cross-sectional area without any assumption regarding the arterial shape using videodensitometry in conjunction with the motion immune dual-energy subtraction technique.

Absolute phasic blood flow measurement in the brain using digital subtraction angiography.

RATIONALE AND OBJECTIVES: In this study, an angiographic method using first-pass distribution analysis was used for quantification of phasic volumetric blood flow. Studies were conducted in an angiographic perfusion phantom and in an animal model (rabbit) after intraarterial injection of contrast material. METHODS: Digital subtraction angiography and first-pass distribution algorithm were used to assess the absolute volumetric flow rates. The method, based on the conservation of contrast material in successive angiographic images, uses the videodensitometric information in the arterial bed. Measurements were made by summing the pixel values in the region of interest, covering the whole perfusion bed. A contrast pass curve was generated for a sequence of images to obtain instantaneous volumetric flow rates. RESULTS: Instantaneous and mean absolute volumetric flow measurements made in the angiographic perfusion phantom and the common carotid artery of the animal models correlated well with validation measurements made using ultrasound flowprobes. The measured (M) and known (K) flow rates in the carotid artery were related by M = 0.87 K + 2.50 mL/minute (r = 0.96, standard error of the estimate = 3.79 mL/minute, n = 25) and M = 0.92 K - 1.00 mL/minute (r = 0.98, standard error of the estimate = 4.04 mL/minute, n = 38) using the videodensitometric and entrance vessel calibration techniques, respectively. Conclusion. Results demonstrate the potential use of the first-pass distribution method in conjunction with digital subtraction angiography for measuring phasic arterial blood flow in vivo.

A digital filtration technique for scatter-glare correction based on thickness estimation.

In order to quantitate anatomical and physiological parameters such as vessel dimensions and volumetric blood flow, it is necessary to make corrections for scatter and veiling glare, which are the major sources of nonlinearities in videodensitometric digital subtraction angiography (DSA). A convolution filtering technique has been investigated to estimate scatter-glare distribution in DSA images without the need to sample the scatter-glare intensity for each patient. This technique utilizes exposure parameters and image gray levels to assign equivalent Lucite thickness for every pixel in the image. The thickness information is then used to estimate scatter-glare intensity on a pixel-by-pixel basis. To test its ability to estimate scatter-glare intensity, the correction technique was applied to images of a Lucite step phantom, anthropomorphic chest phantom, head phantom, and animal models at different thicknesses, projections, and beam energies. The root-mean-square (rms) percentage error of these estimates was obtained by comparison with direct scatter-glare measurements made behind a lead strip. The average rms percentage errors in the scatter-glare estimate for the 25 phantom studies and the 17 animal studies were 6.44% and 7.96%, respectively. These results indicate that the scatter-glare intensity can be estimated with adequate accuracy for a wide range of thicknesses, projections, and beam energies using exposure parameters and gray level information.

CCD camera for dual-energy digital subtraction angiography.

A motion immune dual-energy subtraction technique in which X-ray tube voltage and beam filtration were switched at 30 Hz between 60 kVp (2.0 mm Al filter) and 120 kVp (2.00 mm Al+2.5 mm Cu filter) was previously reported. In this study the effects of camera lag on the dual-energy iodine signal is investigated. The temporal lag of the lead oxide vidicon tested reduced the dual-energy iodine signal by a factor of 2.3, as compared to a mode that included 4 scrub frames between low- and high-energy images, for an iodine phantom with thicknesses of 0-86.0 mg/cm(2), imaged over a 15 cm thick Lucite phantom. On the other hand, the Charge-Coupled Device (CCD) camera has inherently no temporal lag and its versatile scanning characteristics make it near ideal for dual-energy DSA. The CCD camera eliminates the reduction of dual-energy iodine signal, since it does not mix low- and high-energy image data. Another benefit of the CCD camera is that the separation time between low and high-energy images is not limited to the frame period, as is the lead oxide vidicon; and as small as a 5-msec time difference is possible. The short time interval between low and high-energy images minimizes motion misregistration artifacts. Due to these advantages, the CCD camera significantly improves the utility of dual-energy DSA.

Absolute volumetric blood flow measurements using dual-energy digital subtraction angiography.

In recent years, as a solution to the well-documented problems associated with visual interpretation of coronary arteriograms, more physiologic means of assessing coronary artery stenosis are being investigated. Absolute arterial blood flow assessed as a function of time can be a valuable aid in the analysis of functional significance of arterial lesions and obstructions. An absolute volumetric blood flow measurement technique using a motion immune dual-energy subtraction technique is being investigated, where the kVp and filtration are switched at 30 Hz. The low- and high-energy images are corrected for scatter and veiling glare before subtraction. In this technique, the absolute arterial blood flow is calculated by combining the videodensitometric analysis of spatial and temporal aspects concerning the contrast propagation through the arterial bed using tissue suppressed energy subtracted images. The blood flow measurement technique was validated using a pulsatile pump and a flow chamber imaged over a Humanoid chest phantom. A 20-MHz Doppler flow probe was used to validate the measurement of phasic volumetric blood flow. The measured (M) and known (K) mean blood flow for the entrance vessel technique and the videodensitometric calibration technique were related by M = 1.14K - 0.12 ml/s (r2 = 0.98) and M = 1.12K - 0.23 ml/s (r2 = 0.90), respectively. The results indicate that phasic volumetric blood flow can be measured using a CCD camera in conjunction with real time dual-energy subtraction.

Quantification of coronary arterial calcium by dual energy digital subtraction fluoroscopy.

Clinical studies of the heart with fluoroscopy have shown that fluoroscopic visualization of calcium in the coronary arteries is strongly associated with coronary artery disease. However, fluoroscopic detection is limited by its low sensitivity, which is partly due to the interfering background tissue structures and image quantum noise. Moreover, quantification of the absolute amount of calcium in an arterial segment has not been possible. A real-time dual-energy subtraction technique has been investigated as a possible solution to the above problem. In this energy subtraction technique, the kVp and filtration are switched at 30 Hz. In order to assess the potential utility of this videodensitometric technique to quantitate coronary artery calcium, arterial phantoms and excised segments of diseased human arteries were imaged. The low- and high-energy images were corrected for scatter and veiling glare before subtraction. Calcium measurements were made using the tissue-suppressed energy-subtracted images. The estimated calcium phosphate and ashed weights of the calcified arterial segments (N = 20) were highly correlated (slope = 1.04, Intercept = -0.33 mg, r = 0.92).

Absolute cross-sectional area measurements in quantitative coronary arteriography by dual-energy DSA.

Recent studies have emphasized the limitations of conventional coronary angiography. These limitations include the lack of correlation between the severity of coronary stenosis as estimated from coronary angiograms and the actual severity of stenotic lesions measured in postmortem hearts. As a result, attempts have been made to quantitate luminal dimension more precisely. The application of quantitative digital subtraction angiography (DSA) in the assessment of coronary artery lesion dimension has been limited by cardiac and respiratory motion artifacts. We have reported previously on a motion-immune dual-energy (DE) cardiac mode in which kVp and filtration are switched at 30 Hz. To assess the potential advantages of a videodensitometric technique for quantification of absolute vessel cross-sectional area (CSA), three different quantitative coronary arteriography (QCA) algorithms were compared. The three algorithms under comparison were a videodensitometric (V) algorithm, which does not require any geometric assumption for absolute vessel CSA measurement, and videodensitometric (VC) and edge detection (ED) algorithms, which do require the assumption of circular cross-section for CSA measurements. A cylindrical vessel phantom (0.5-4.75 mm in diameter) and a crescentic vessel phantom, producing 25% to 90% area stenosis, were imaged over the chest of a humanoid phantom. The low- and high-energy images were corrected for scatter and veiling glare before energy subtraction. For CSA measurements in crescentic vessel phantoms, the V algorithm produced significantly improved results (slope = 0.87, intercept = 0.51 mm2, r = .95) when compared to the VC (slope = 1.05, intercept = 4.19 mm2, r = .75) and the ED (slope = 1.57, intercept = 5.21 mm2, r = .60) algorithms.