Accelerated free-breathing liver fat and R 2 * quantification using multi-echo stack-of-radial MRI with motion-resolved multidimensional regularized reconstruction: Initial retrospective evaluation.

PURPOSE: To improve image quality, mitigate quantification biases and variations for free-breathing liver proton density fat fraction (PDFF) and R 2 * $$ {\mathrm{R}}_2^{\ast } $$ quantification accelerated by radial k-space undersampling. METHODS: A free-breathing multi-echo stack-of-radial MRI method was developed with compressed sensing with multidimensional regularization. It was validated in motion phantoms with reference acquisitions without motion and in 11 subjects (6 patients with nonalcoholic fatty liver disease) with reference breath-hold Cartesian acquisitions. Images, PDFF, and R 2 * $$ {\mathrm{R}}_2^{\ast } $$ maps were reconstructed using different radial view k-space sampling factors and reconstruction settings. Results were compared with reference-standard results using Bland-Altman analysis. Using linear mixed-effects model fitting (p < 0.05 considered significant), mean and SD were evaluated for biases and variations of PDFF and R 2 * $$ {\mathrm{R}}_2^{\ast } $$ , respectively, and coefficient of variation on the first echo image was evaluated as a surrogate for image quality. RESULTS: Using the empirically determined optimal sampling factor of 0.25 in the accelerated in vivo protocols, mean differences and limits of agreement for the proposed method were [-0.5; -33.6, 32.7] s-1 for R 2 * $$ {\mathrm{R}}_2^{\ast } $$ and [-1.0%; -5.8%, 3.8%] for PDFF, close to those of a previous self-gating method using fully sampled radial views: [-0.1; -27.1, 27.0] s-1 for R 2 * $$ {\mathrm{R}}_2^{\ast } $$ and [-0.4%; -4.5%, 3.7%] for PDFF. The proposed method had significantly lower coefficient of variation than other methods (p < 0.001). Effective acquisition time of 64 s or 59 s was achieved, compared with 171 s or 153 s for two baseline protocols with different radial views corresponding to sampling factor of 1.0. CONCLUSION: This proposed method may allow accelerated free-breathing liver PDFF and R 2 * $$ {\mathrm{R}}_2^{\ast } $$ mapping with reduced biases and variations.

Ferumoxytol-Enhanced Cardiac Cine MRI Reconstruction Using a Variable-Splitting Spatiotemporal Network.

BACKGROUND: Balanced steady-state free precession (bSSFP) imaging is commonly used in cardiac cine MRI but prone to image artifacts. Ferumoxytol-enhanced (FE) gradient echo (GRE) has been proposed as an alternative. Utilizing the abundance of bSSFP images to develop a computationally efficient network that is applicable to FE GRE cine would benefit future network development. PURPOSE: To develop a variable-splitting spatiotemporal network (VSNet) for image reconstruction, trained on bSSFP cine images and applicable to FE GRE cine images. STUDY TYPE: Retrospective and prospective. SUBJECTS: 41 patients (26 female, 53 ± 19 y/o) for network training, 31 patients (19 female, 49 ± 17 y/o) and 5 healthy subjects (5 female, 30 ± 7 y/o) for testing. FIELD STRENGTH/SEQUENCE: 1.5T and 3T, bSSFP and GRE. ASSESSMENT: VSNet was compared to VSNet with total variation loss, compressed sensing and low rank methods for 14× accelerated data. The GRAPPA×2/×3 images served as the reference. Peak signal-to-noise-ratio (PSNR), structural similarity index (SSIM), left ventricular (LV) and right ventricular (RV) end-diastolic volume (EDV), end-systolic volume (ESV), and ejection fraction (EF) were measured. Qualitative image ranking and scoring were independently performed by three readers. Latent scores were calculated based on scores of each method relative to the reference. STATISTICS: Linear mixed-effects regression, Tukey method, Fleiss' Kappa, Bland-Altman analysis, and Bayesian categorical cumulative probit model. A P-value <0.05 was considered statistically significant. RESULTS: VSNet achieved significantly higher PSNR (32.7 ± 0.2), SSIM (0.880 ± 0.004), rank (2.14 ± 0.06), and latent scores (-1.72 ± 0.22) compared to other methods (rank >2.90, latent score < -2.63). Fleiss' Kappa was 0.52 for scoring and 0.61 for ranking. VSNet showed no significantly different LV and RV ESV (P = 0.938) and EF (P = 0.143) measurements, but statistically significant different (2.62 mL) EDV measurements compared to the reference. CONCLUSION: VSNet produced the highest image quality and the most accurate functional measurements for FE GRE cine images among the tested 14× accelerated reconstruction methods. LEVEL OF EVIDENCE: 3 TECHNICAL EFFICACY: Stage 1.

Has the concept of polyspermy prevention been invented in the laboratory?

There is no evidence, nor need, for a fast block to polyspermy in animal oocytes. The idea that oocytes have evolved a mechanism to allow the entry of one spermatozoon and repel all others has, however, gained consensus over the last century. The main culprit is the sea urchin, which has been used for over a century in in vitro studies of the fertilization process. Images of sea urchin oocytes with thousands of sperm attached to the surface are commonplace in textbooks and appeal to the nature of the reader implying an intriguing surface mechanism of sperm selection despite these oocytes being fixed for photography (Figure ). The abundance of gametes in this marine invertebrate and the ease of experimentation have given us the possibility to elucidate many aspects of the mechanism of fertilization, but has also led to ongoing controversies in reproductive biology, one being polyspermy prevention. Kinetic experiments by Rothschild and colleagues in the 1950s led to the hypothesis of a fast partial block to polyspermy in sea urchin oocytes that reduced the probability of a second spermatozoon from entering the oocyte by 1/20th. In the 1970s, Jaffe and colleagues suggested, with circumstantial evidence, that this partial block was due to the sperm-induced depolarization of the oocyte plasma membrane. However, the fate of supernumerary spermatozoa is determined well before the plasma membrane of the oocyte depolarizes. Transmembrane voltage does not serve to regulate sperm entry. Scholastic texts have inadvertently promulgated this concept across the animal kingdom with no logical correlation or experimentation and, as of today, a molecular mechanism to regulate sperm entry in oocytes has not been identified.

Platelets from patients with myeloproliferative neoplasms have increased numbers of mitochondria that are hypersensitive to depolarization by thrombin.

Thrombosis is one of the cardinal manifestations of myeloproliferative neoplasms (MPN). The mechanisms leading to a prothrombotic state in MPN are complex and remain poorly understood. Platelet mitochondria play a role in platelet activation, but their number and function have not been extensively explored in MPN to date. We observed an increased number of mitochondria in platelets from MPN patients compared with healthy donors. MPN patients had an increased proportion of dysfunctional platelet mitochondria. The fraction of platelets with depolarized mitochondria at rest was increased in essential thrombocythemia (ET) patients and the mitochondria were hypersensitive to depolarization following thrombin agonist stimulation. Live microscopy showed a stochastic process in which a higher proportion of individual ET platelets underwent mitochondrial depolarization and after a shorter agonist exposure compared to healthy donors. Depolarization was immediately followed by ballooning of the platelet membrane, which is a feature of procoagulant platelets. We also noted that the mitochondria of MPN patients were on average located nearer the platelet surface and we observed extrusion of mitochondria from the platelet surface as microparticles. These data implicate platelet mitochondria in a number of prothrombotic phenomena. Further studies are warranted to assess whether these findings correlate with clinical thrombotic events.

Male Germ Cell Telomeres and Chemical Pollutants.

In recent decades, male infertility has been correlated with the shortening of sperm telomeres. Telomeres regulate the reproductive lifespan by mediating the synapsis and homologous recombination of chromosomes during gametogenesis. They are composed of thousands of hexanucleotide DNA repeats (TTAGGG) that are coupled to specialized shelterin complex proteins and non-coding RNAs. Telomerase activity in male germ cells ensures that the telomere length is maintained at maximum levels during spermatogenesis, despite telomere shortening due to DNA replication or other genotoxic factors such as environmental pollutants. An emerging body of evidence has associated an exposure to pollutants with male infertility. Although telomeric DNA may be one of the important targets of environmental pollutants, only a few authors have considered it as a conventional parameter for sperm function. The aim of this review is to provide comprehensive and up-to-date data on the research carried out so far on the structure/function of telomeres in spermatogenesis and the influence of environmental pollutants on their functionality. The link between pollutant-induced oxidative stress and telomere length in germ cells is discussed.

Undersampling artifact reduction for free-breathing 3D stack-of-radial MRI based on a deep adversarial learning network.

PURPOSE: Stack-of-radial MRI allows free-breathing abdominal scans, however, it requires relatively long acquisition time. Undersampling reduces scan time but can cause streaking artifacts and degrade image quality. This study developed deep learning networks with adversarial loss and evaluated the performance of reducing streaking artifacts and preserving perceptual image sharpness. METHODS: A 3D generative adversarial network (GAN) was developed for reducing streaking artifacts in stack-of-radial abdominal scans. Training and validation datasets were self-gated to 5 respiratory states to reduce motion artifacts and to effectively augment the data. The network used a combination of three loss functions to constrain the anatomy and preserve image quality: adversarial loss, mean-squared-error loss and structural similarity index loss. The performance of the network was investigated for 3-5 times undersampled data from 2 institutions. The performance of the GAN for 5 times accelerated images was compared with a 3D U-Net and evaluated using quantitative NMSE, SSIM and region of interest (ROI) measurements as well as qualitative scores of radiologists. RESULTS: The 3D GAN showed similar NMSE (0.0657 vs. 0.0559, p = 0.5217) and significantly higher SSIM (0.841 vs. 0.798, p < 0.0001) compared to U-Net. ROI analysis showed GAN removed streaks in both the background air and the tissue and was not significantly different from the reference mean and variations. Radiologists' scores showed GAN had a significant improvement of 1.6 point (p = 0.004) on a 4-point scale in streaking score while no significant difference in sharpness score compared to the input. CONCLUSION: 3D GAN removes streaking artifacts and preserves perceptual image details.

Comparison of sperm preparation methods to improve the recovery of mature spermatozoa in sub-fertile males.

The integrity of chromatin in the spermatozoon is essential for reproductive outcome. The aim of this study was to evaluate the most effective and cost-effective method to reduce the percentage of spermatozoa with defects in chromatin decondensation for use in assisted reproductive technologies (ART) procedures. Sperm samples from 15 sub-fertile males were examined at CFA Naples to determine the sperm decondensation index (SDI), using the aniline blue test, before and after preparation, comparing density gradients with two different swim-up approaches. All three techniques led to a reduction in decondensed spermatozoa with no statistical difference (P > 0.05) between the control and the treated sperm. In contrast, we found a highly significant decrease in SDI (P < 0.01) after the two swim-up methods in all the samples, confirming the efficacy of these methods in lowering the percentage of chromatin compaction damage. There was no statistical difference between the two swim-up methods, however swim-up from the pellet led to improved count, motility and the percentage of normal condensed spermatozoa. We suggest that swim-up from the pellet be used in ART on sub-fertile males, both to reduce cell stress by multiple centrifugation and improve the recovery rate of mature spermatozoa.

Can sperm quality influence embryo development and its ploidy? Analysis of 811 blastocysts obtained from different sperm sources.

The aim of our study was to evaluate the correlation between sperm quality and ploidy status of the derived blastocysts. We performed a retrospective analysis on a restricted pool of patients enrolling only those who had no female factors. Male patients with genetic factors affecting spermatogenesis were also excluded. We chose a maternal age ≤38 years to decrease the female factor, therefore the male factor was the main component of sterility. We divided the patients in four groups based on semen quality and comparing fertilization, pregnancy and euploidy rates above all. In total, 201 intracytoplasmic sperm injection (ICSI) cycles were enrolled in the study. Cycles were divided into four groups, according to semen source: normal semen, oligoasthenoteratozoospermia (OAT), cryptospermia or non-obstructive azoospermia (NOA). An extremely statistically lower fertilization rate was found in NOA patients. Unexpectedly, no differences were detected in blastocyst formation, euploidy, aneuploidy and mosaicism rates among the four groups. Interestingly, we also found a higher abortion rate comparing NOA to normal semen with an odds ratio of 4.67. In our study no statistically significant differences among the analyzed groups were found, showing little or no effect at all using spermatozoa from different semen sources or quality. This may be linked to the oocyte competence of fixing sperm DNA damage and it could be hypothesized that only sperm with a good rate of DNA integrity are able to fertilize the oocyte, explaining why poor quality semen is reflected in a low fertilization rate without effect on ploidy.

Accelerated k-space shift calibration for free-breathing stack-of-radial MRI quantification of liver fat and R2∗.

PURPOSE: To develop an accelerated k-space shift calibration method for free-breathing 3D stack-of-radial MRI quantification of liver proton-density fat fraction (PDFF) and R2∗ . METHODS: Accelerated k-space shift calibration was developed to partially skip acquisition of k-space shift data in the through-plane direction then interpolate in processing, as well as to reduce the in-plane averages. A multi-echo stack-of-radial sequence with the baseline calibration was evaluated on a phantom versus vendor-provided reference-standard PDFF and R2∗ values at 1.5T, and in 13 healthy subjects and 5 clinical subjects at 3T with respect to reference-standard breath-hold Cartesian acquisitions. PDFF and R2∗ maps were calculated with different calibration acceleration factors offline and compared to reference-standard values using Bland-Altman analysis. Bias and uncertainty were evaluated using normal distribution and Bayesian probability of difference (P < .05 considered significant). RESULTS: Bland-Altman plots of phantom and in vivo data showed that substantial acceleration was highly feasible in both through-plane and in-plane directions. Compared to the baseline calibration without acceleration, Bayesian analysis revealed no significant differences on biases and uncertainties of PDFF and R2∗ measurements with all acceleration methods in this study, except the method with through-plane acceleration equaling slices and averages equaling 20 for PDFF and R2∗ (both P < .001) for the phantom. A six-fold reduction in equivalent calibration acquisition time (time saving ≥25 s and ≥80.7%) was achieved using recommended acceleration factors for the in vivo protocols in this study. CONCLUSION: This proposed method may allow accelerated calibration for free-breathing stack-of-radial MRI PDFF and R2∗ mapping.

Modulating oxidative stress and epigenetic homeostasis in preimplantation IVF embryos.

Assisted reproductive technology is today considered a safe and reliable medical intervention, with healthy live births a reality for many IVF and ICSI treatment cycles. However, there are increasing numbers of published reports describing epigenetic/imprinting anomalies in children born as a result of these procedures. These anomalies have been attributed to methylation errors in embryo chromatin remodelling during in vitro culture. Here we re-visit three concepts: (1) the so-called 'in vitro toxicity' of 'essential amino acids' before the maternal to zygotic transition period; (2) the effect of hyperstimulation (controlled ovarian hyperstimulation) on homocysteine in the oocyte environment and the effect on methylation in the absence of essential amino acids; and (3) the fact/postulate that during the early stages of development the embryo undergoes a 'global' demethylation. Methylation processes require efficient protection against oxidative stress, which jeopardizes the correct acquisition of methylation marks as well as subsequent methylation maintenance. The universal precursor of methylation [by S-adenosyl methionine (SAM)], methionine, 'an essential amino acid', should be present in the culture. Polyamines, regulators of methylation, require SAM and arginine for their syntheses. Cystine, another 'semi-essential amino acid', is the precursor of the universal protective antioxidant molecule: glutathione. It protects methylation marks against some undue DNA demethylation processes through ten-eleven translocation (TET), after formation of hydroxymethyl cytosine. Early embryos are unable to convert homocysteine to cysteine as the cystathionine ß-synthase pathway is not active. In this way, cysteine is a 'real essential amino acid'. Most IVF culture medium do not maintain methylation/epigenetic processes, even in mouse assays. Essential amino acids should be present in human IVF medium to maintain adequate epigenetic marking in preimplantation embryos. Furthermore, morphological and morphometric data need to be re-evaluated, taking into account the basic biochemical processes involved in early life.

Free-breathing 3D stack-of-radial MRI quantification of liver fat and R2* in adults with fatty liver disease.

PURPOSE: To investigate the agreement, intra-session repeatability, and inter-reader agreement of liver proton-density fat fraction (PDFF) and R2* quantification using free-breathing 3D stack-of-radial MRI, with and without self-gated motion compensation, compared to reference breath-hold techniques in subjects with fatty liver disease (FLD). METHODS: In this institutional review board-approved prospective study, thirty-eight adults with FLD and/or iron overload (24 male, 58 ± 12 years) were imaged at 3T using free-breathing stack-of-radial MRI, breath-hold 3D Cartesian MRI, and breath-hold single-voxel MR spectroscopy (SVS). Each sequence was acquired twice in random order. To assess agreement compared to reference breath-hold techniques, the dependency of liver PDFF and/or R2* quantification on the sequence, radial sampling factor, and radial self-gating temporal resolution was assessed by calculating the Bayesian mean difference (MDB) of the posteriors. Intra-session repeatability and inter-reader agreement (two independent readers) were assessed by the coefficient of repeatability (CR) and intraclass correlation coefficient (ICC), respectively. RESULTS: Thirty-five participants (21 male, 57 ± 12 years) were included for analysis. Both free-breathing radial MRI techniques (with and without self-gating) achieved ICC ≥ 0.92 for quantifying PDFF and R2*, and quantified PDFF with MDB < 1.2% compared to breath-hold techniques. Free-breathing radial MRI required self-gating to accurately quantify R2* (MDB < 10s-1 with self-gating; MDB < 50s-1 without self-gating). The radial sampling factor affected PDFF and R2* quantification while the radial self-gating temporal resolution only affected R2* quantification. Repeated self-gated free-breathing radial MRI scans achieved CR < 3% and CR < 27 s-1 for PDFF and R2*, respectively. CONCLUSION: A free-breathing stack-of-radial MRI technique with self-gating demonstrated agreement, repeatability, and inter-reader agreement compared to reference breath-hold techniques for quantification of liver PDFF and R2* in adults with FLD.

CD36 promotes vasculogenic mimicry in melanoma by mediating adhesion to the extracellular matrix.

BACKGROUND: The formation of blood vessels within solid tumors directly contributes to cancer growth and metastasis. Until recently, tumor vasculature was thought to occur exclusively via endothelial cell (EC) lined structures (i.e. angiogenesis), but a second source of tumor vasculature arises from the cancer cells themselves, a process known as vasculogenic mimicry (VM). While it is generally understood that the function of VM vessels is the same as that of EC-lined vessels (i.e. to supply oxygen and nutrients to the proliferating cancer cells), the molecular mechanisms underpinning VM are yet to be fully elucidated. METHODS: Human VM-competent melanoma cell lines were examined for their VM potential using the in vitro angiogenesis assays (Matrigel), together with inhibition studies using small interfering RNA and blocking monoclonal antibodies. Invasion assays and adhesion assays were used to examine cancer cell function. RESULTS: Herein we demonstrate that CD36, a cell surface glycoprotein known to promote angiogenesis by ECs, also supports VM formation by human melanoma cancer cells. In silico analysis of CD36 expression within the melanoma cohort of The Cancer Genome Atlas suggests that melanoma patients with high expression of CD36 have a poorer clinical outcome. Using in vitro 'angiogenesis' assays and CD36-knockdown approaches, we reveal that CD36 supports VM formation by human melanoma cells as well as adhesion to, and invasion through, a cancer derived extracellular matrix substrate. Interestingly, thrombospondin-1 (TSP-1), a ligand for CD36 on ECs that inhibits angiogenesis, has no effect on VM formation. Further investigation revealed a role for laminin, but not collagen or fibronectin, as ligands for CD36 expressing melanoma cells. CONCLUSIONS: Taken together, this study suggests that CD36 is a novel regulator of VM by melanoma cancer cells that is facilitated, at least in part, via integrin-α3 and laminin. Unlike angiogenesis, VM is not perturbed by the presence of TSP-1, thus providing new information on differences between these two processes of tumor vascularization which may be exploited to combat cancer progression.

MTHFR (methylenetetrahydrofolate reductase: EC 1.5.1.20) SNPs (single-nucleotide polymorphisms) and homocysteine in patients referred for investigation of fertility.

PURPOSE: MTHFR, one of the major enzymes in the folate cycle, is known to acquire single-nucleotide polymorphisms that significantly reduce its activity, resulting in an increase in circulating homocysteine. Methylation processes are of crucial importance in gametogenesis, involved in the regulation of imprinting and epigenetic tags on DNA and histones. We have retrospectively assessed the prevalence of MTHFR SNPs in a population consulting for infertility according to gender and studied the impact of the mutations on circulating homocysteine levels. METHODS: More than 2900 patients having suffered at least two miscarriages (2 to 9) or two failed IVF/ICSI (2 to 10) attempts were included for analysis of MTHFR SNPs C677T and A1298C. Serum homocysteine levels were measured simultaneously. RESULTS: We observed no difference in the prevalence of different genetic backgrounds between men and women; only 15% of the patients were found to be wild type. More than 40% of the patients are either homozygous for one SNP or compound heterozygous carriers. As expected, the C677T SNP shows the greatest adverse effect on homocysteine accumulation. The impact of MTHFR SNPs on circulating homocysteine is different in men than in women. CONCLUSIONS: Determination of MTHFR SNPs in both men and women must be seriously advocated in the presence of long-standing infertility; male gametes, from MTHFR SNPs carriers, are not exempted from exerting a hazardous impact on fertility. Patients should be informed of the pleiotropic medical implications of these SNPs for their own health, as well as for the health of future children.

Free-Breathing Volumetric Liver R2* and Proton Density Fat Fraction Quantification in Pediatric Patients Using Stack-of-Radial MRI With Self-Gating Motion Compensation.

BACKGROUND: Stack-of-radial multiecho gradient-echo MRI is promising for free-breathing liver R2* quantification and may benefit children. PURPOSE: To validate stack-of-radial MRI with self-gating motion compensation in phantoms, and to evaluate it in children. STUDY TYPE: Prospective. PHANTOMS: Four vials with different R2* driven by a motion stage. SUBJECTS: Sixteen pediatric patients with suspected nonalcoholic fatty liver disease or steatohepatitis (five females, 13 ± 4 years, body mass index 29.2 ± 8.6 kg/m2 ). FIELD STRENGTH/SEQUENCES: Stack-of-radial, and 2D and 3D Cartesian multiecho gradient-echo sequences at 3T. ASSESSMENT: Ungated and gated stack-of-radial proton density fat fraction (PDFF) and R2* maps were reconstructed without and with self-gating motion compensation. Stack-of-radial R2* measurements of phantoms without and with motion were validated against reference 2D Cartesian results of phantoms without motion. In subjects, free-breathing stack-of-radial and reference breath-hold 3D Cartesian were acquired. Subject inclusion for statistical analysis and region of interest placement were determined independently by two observers. STATISTICAL TESTS: Phantom results were fitted with a weighted linear model. Demographic differences between excluded and included subjects were tested by multivariate analysis of variance. PDFF and R2* measurements were compared using Bland-Altman analysis. Interobserver agreement was assessed by the intraclass correlation coefficient (ICC). RESULTS: Ungated stack-of-radial R2* inside moving phantom vials showed a significant positive bias of 64.3 s-1 (P < 0.00001), unlike gated results (P > 0.31). Subject inclusion decisions for statistical analysis from two observers were consistent. No significant differences were found between four excluded and 12 included subjects (P = 0.14). Compared to breath-hold Cartesian, ungated and gated free-breathing stack-of-radial exhibited mean R2* differences of 18.5 s-1 and 3.6 s-1 . Mean PDFF differences were 1.1% and 1.0% for ungated and gated measurements, respectively. Interobserver agreement was excellent (ICC for PDFF = 0.99, ICC for R2* = 0.90; P < 0.0003). DATA CONCLUSION: Stack-of-radial MRI with self-gating motion compensation seems to allow free-breathing liver R2* and PDFF quantification in children. LEVEL OF EVIDENCE: 2 TECHNICAL EFFICACY STAGE: 2.

Platelets disrupt vasculogenic mimicry by cancer cells.

Tumour vasculature supports the growth and progression of solid cancers with both angiogenesis (endothelial cell proliferation) and vasculogenic mimicry (VM, the formation of vascular structures by cancer cells themselves) predictors of poor patient outcomes. Increased circulating platelet counts also predict poor outcome for cancer patients but the influence of platelets on tumour vasculature is incompletely understood. Herein, we show with in vitro assays that platelets did not influence angiogenesis but did actively inhibit VM formation by cancer cell lines. Both platelet sized beads and the releasates from platelets were partially effective at inhibiting VM formation suggesting that direct contact maximises the effect. Platelets also promoted cancer cell invasion in vitro. B16F10 melanomas in Bcl-xPlt20/Plt20 thrombocytopenic mice showed a higher content of VM than their wildtype counterparts while angiogenesis did not differ. In a xenograft mouse model of breast cancer with low-dose aspirin to inactivate the platelets, the burden of MDA-MB-231-LM2 breast cancer cells was reduced and the gene expression profile of the cancer cells was altered; but no effect on tumour vasculature was observed. Taken together, this study provides new insights into the action of platelets on VM formation and their involvement in cancer progression.

DAAM1 and PREP are involved in human spermatogenesis.

During differentiation of the male gamete, there is a massive remodelling in the shape and architecture of all the cells in the seminiferous epithelium. The cytoskeleton, as well as many associated proteins, plays a pivotal role in this process. To better characterise the factors involved, we analysed two proteins: the formin, dishevelled-associated activator of morphogenesis 1 (DAAM1), which participates in the regulation of actin polymerisation, and the protease, prolyl endopeptidase (PREP), engaged in microtubule-associated processes. In our previous studies we demonstrated their involvement in cytoskeletal dynamics necessary for correct postnatal development of the rat testis. Here, we used samples of testicular tissue obtained from infertile men by testicular sperm extraction and the spermatozoa of asthenoteratozoospermic patients. By western blot and immunofluorescent analysis, we found that DAAM1 and PREP expression and localisation were impaired in both the testis and spermatozoa, and in particular in the midpiece as well as in the principal and end-pieces of the flagella, as compared with spermatozoa of normospermic men. Our results provide new knowledge of the dynamics of spermatogenesis, raising the possibility of using DAAM1 and PREP as new markers of normal fertility.

Effect of respiratory motion on free-breathing 3D stack-of-radial liver R2∗ relaxometry and improved quantification accuracy using self-gating.

PURPOSE: To develop an accurate free-breathing 3D liver R2∗ mapping approach and to evaluate it in vivo. METHODS: A free-breathing multi-echo stack-of-radial sequence was applied in 5 normal subjects and 6 patients at 3 Tesla. Respiratory motion compensation was implemented using the inherent self-gating signal. A breath-hold Cartesian acquisition was the reference standard. Proton density fat fraction and R2∗ were measured and compared between radial and Cartesian methods using Bland-Altman plots. The normal subject results were fitted to a linear mixed model (P < .05 considered significant). RESULTS: Free-breathing stack-of-radial without self-gating exhibited signal attenuation in echo images and artifactually elevated apparent R2∗ values. In the Bland-Altman plots of normal subjects, compared to breath-hold Cartesian, free-breathing stack-of-radial acquisitions of 22, 30, 36, and 44 slices, had mean R2∗ differences of 27.4, 19.4, 10.9, and 14.7 s-1 with 800 radial views, and they had 18.4, 11.9, 9.7, and 27.7 s-1 with 404 views, which were reduced to 0.4, 0.9, -0.2, and -0.7 s-1 and to -1.7, -1.9, -2.1, and 0.5 s-1 with self-gating, respectively. No substantial proton density fat fraction differences were found. The linear mixed model showed free-breathing radial R2∗ results without self-gating were significantly biased by 17.2 s-1 averagely (P = .002), which was eliminated with self-gating (P = .930). Proton density fat fraction results were not different (P > .234). For patients, Bland-Altman plots exhibited mean R2∗ differences of 14.4 and 0.1 s-1 for free-breathing stack-of-radial without self-gating and with self-gating, respectively, but no substantial proton density fat fraction differences. CONCLUSION: The proposed self-gating method corrects the respiratory motion bias and enables accurate free-breathing stack-of-radial quantification of liver R2∗ .

Improved accuracy of apparent diffusion coefficient quantification using a fully automatic noise bias compensation method: Preliminary evaluation in prostate diffusion weighted imaging.

Noise in diffusion magnetic resonance imaging can introduce bias in apparent diffusion coefficient (ADC) quantification. Previous studies proposed methods that are site-specific techniques as research tools with limited availability and typically require manual intervention, not completely ready to use in the clinical environment. The purpose of this study was to develop a fully automatic computational method to correct noise bias in ADC quantification and perform a preliminary evaluation in the clinical prostate diffusion weighted imaging (DWI). Using a pseudo replica approach for the noise map calculation as well as a direct mapping and a stepwise Chebychev polynomial modelling approach for the ADC fitting, a fully automatic noise-bias-compensated ADC calculation method was proposed and implemented both on the scanner and offline. The proposed method was validated in a computer simulation and a standardized diffusion phantom with ground-truth values. Two in vivo studies were performed to evaluate the proposed method in the clinical environment. The first in vivo study performed acquisitions using a clinically routine prostate DWI protocol on 29 subjects to evaluate the consistency between simulated and empirical results. In the second in vivo study, prostate ADC values of 14 subjects were compared between data acquired with external coils only and reconstructed with the proposed method vs. acquired with external combined with endorectal coils and reconstructed with the conventional method. In statistical analyses, p < 0.05 was regarded as significantly different. In the computer simulation, the proposed method showed smaller error percentage than the other methods and was significantly different (p < 2.2 × 10-16). With low signal-to-noise ratio (SNR), the conventional method underestimated ADC values compared to the ground truth values of the diffusion phantom, while the results of the proposed method were more consistent with the ground truth values. Statistical analyses showed no significant differences between measured and simulated results in the first in vivo study (p = 0.5618). Data from the second in vivo study showed that agreement between ADC measured with external coils only and combined coils was improved for the proposed method (mean bias: 0.04 × 10-3 mm2/s, 95% confidence interval (CI) = [-0.01, 0.09] × 10-3 mm2/s, p = 0.187), compared to the conventional method (mean bias: -0.12 × 10-3 mm2/s, 95% CI = [-0.17, -0.06] × 10-3 mm2/s, p < 0.0001). The proposed method compensates noise bias in low-SNR diffusion-weighted acquisitions and results show improved ADC quantification accuracy in the prostate. This method may be suitable for both clinical imaging and research utilizing ADC quantification.

DNA Methylation Patterns in the Early Human Embryo and the Epigenetic/Imprinting Problems: A Plea for a More Careful Approach to Human Assisted Reproductive Technology (ART).

An increasing number of publications indicate that babies born after IVF (in vitro fertilization) procedures have higher rates of anomalies related to imprinting/epigenetic changes, which may be attributed to suboptimal culture conditions. Appropriate maintenance of DNA methylation during the first few days of an in vitro culture requires a supply of methyl donors, which are lacking in current in vitro culture systems. The absence of protection against oxidative stress in the culture increases the risks for errors in methylation. A decrease in the methylation processes is sometimes observed immediately post fertilization, due to delays that occur during the maternal⁻zygotic transition period. Care should be exercised in ART (assisted reproductive technology) procedures in order to avoid the risk of generating errors in methylation during the in vitro culture period immediately post fertilization, which has an impact on imprinting/epigenetics. Formulation of IVF culture media needs to be re-assessed in the perspective of current knowledge regarding embryo physiology.