Identification of neuroactive steroids and their precursors and metabolites in adult male rat brain

Steroids in the brain arise both from local synthesis and from peripheral sources and have a variety of effects on neuronal function. However, there is little direct chemical evidence for the range of steroids present in brain or of the pathways for their synthesis and inactivation. This information is a prerequisite for understanding the regulation and function of brain steroids. After extraction from adult male rat brain, we have fractionated free steroids and their sulfate esters and then converted them to heptafluorobutyrate or methyloxime-trimethylsilyl ether derivatives for unequivocal identification and assay by gas chromatography analysis and selected ion monitoring mass spectrometry. In the free steroid fraction, corticosterone, 3 alpha, 5 alpha-tetrahydrodeoxycorticosterone, testosterone, and dehydroepiandrosterone were found in the absence of detectable precursors usually found in endocrine glands, indicating peripheral sources and/or alternative synthetic pathways in brain. Conversely, the potent neuroactive steroid 3 alpha, 5 alpha-tetrahydroprogesterone ( allopregnanolone) was found in the presence of its precursors pregnenolone, progesterone, and 5 alpha-dihydroprogesterone. Furthermore, the presence of 3 alpha-, 11 alpha-, 17 alpha-, and 20 alpha-hydroxylated metabolites of 3 alpha, 5 alpha-tetrahydroprogesterone implicated possible inactivation pathways for this steroid. The 20 alpha-reduced metabolites could also be found for pregnenolone, progesterone, and 5 alpha-dihydroprogesterone, introducing a possible regulatory diversion from the production of 3 alpha, 5 alpha-tetrahydroprogesterone. In the steroid sulfate fraction, dehydroepiandrostrone sulfate was identified but not pregnenolone sulfate. Although pharmacologically active, identification of the latter appears to be an earlier methodological artifact, and the compound is thus of doubtful physiological significance in the adult brain. Our results provide a basis for elucidating the origins and regulation of brain steroids

Volumetric reconstruction from printed films: Enabling 30 year longitudinal analysis in MR neuroimaging

Hospitals often hold historical MR image data printed on films without being able to make it accessible to modern image processing techniques. Having the possibility to recover geometrically consistent, volumetric images from scans acquired decades ago will enable more comprehensive, longitudinal studies to understand disease progressions. In this paper, we propose a consistent framework to reconstruct a volumetric representation from printed films holding thick single-slice brain MR image acquisitions dating back to the 1980's. We introduce a flexible framework based on semi-automatic slice extraction, followed by automated slice-to-volume registration with inter-slice transformation regularisation and slice intensity correction. Our algorithm is robust against numerous detrimental effects being present in archaic films. A subsequent, isotropic total variation deconvolution technique revitalises the visual appearance of the obtained volumes. We assess the accuracy and perform the validation of our reconstruction framework on a uniquely long-term MRI dataset where a ground-truth is available. This method will be used to facilitate a robust longitudinal analysis spanning 30 years of MRI scans

A thirty year clinical and MRI observational study of multiple sclerosis and clinically isolated syndromes

OBJECTIVE: Clinical outcomes in multiple sclerosis (MS) are highly variable. We aim to determine the long-term clinical outcomes in MS, and to identify early prognostic features of these outcomes. METHODS: 132 people presenting with a clinically isolated syndrome (CIS) were prospectively recruited between 1984-87, and followed up clinically and radiologically 1, 5, 10, 14, 20 and now 30 years later. All available notes and magnetic resonance imaging (MRI) scans were reviewed, and MS was defined according to the 2010 McDonald criteria. RESULTS: Clinical outcome data was obtained in 120 participants at 30 years. Eighty were known to have developed MS by 30 years. Expanded disability status scale (EDSS) scores were available in 107 participants, of whom 77 had MS: thirty-two (42%) remained fully ambulatory (EDSS ≤3.5) all of whom had relapsing-remitting MS (RRMS), three (4%) had RRMS and EDSS >3.5, 26 (34%) had secondary progressive MS (all had EDSS >3.5), and MS contributed to death in 16 (20%). Of those with MS, 11 have been treated with a DMT. The strongest early predictors (within 5 years of presentation) of secondary progressive MS (SPMS) at 30 years were presence of baseline infratentorial lesions and deep white matter lesions at one year. INTERPRETATION: Thirty years after onset, in a largely untreated cohort, there was a divergence of MS outcomes; some people accrued substantial disability early on, while others ran a more favourable long-term course. These outcomes could, in part, be predicted by radiological findings from within a year of first presentation

Joint Multimodal Segmentation of Clinical CT and MR from Hip Arthroplasty Patients

Magnetic resonance imaging (MRI) is routinely employed to assess muscular response and presence of inflammatory reactions in patients treated with metal-on-metal hip arthroplasty, driving the decision for revision surgery. However, MRI is lacking contrast for bony structures and as a result orthopaedic surgical planning is mostly performed on computed tomography images. In this paper, we combine the complementary information of both modalities into a novel framework for the joint segmentation of healthy and pathological musculoskeletal structures as well as implants on all images. Our processing pipeline is fully automated and was designed to handle the highly anisotropic resolution of clinical MR images by means of super resolution reconstruction. The accuracy of the intra-subject multimodal registration was improved by employing a non-linear registration algorithm with hard constraints on the deformation of bony structures, while a multi-atlas segmentation propagation approach provided robustness to the large shape variability in the population. The suggested framework was evaluated in a leave-one-out cross-validation study on 20 hip sides. The proposed pipeline has potential for the extraction of clinically relevant imaging biomarkers for implant failure detection

Impact of GnRH analogues on oocyte/embryo quality and embryo development in in vitro fertilization/intracytoplasmic sperm injection cycles: a case control study

<p>Abstract</p> <p>Background</p> <p>Despite the clinical outcomes of ovarian stimulation with either GnRH-agonist or GnRH-antagonist analogues for in vitro fertilization (IVF) being well analysed, the effect of analogues on oocyte/embryo quality and embryo development is still not known in detail. The aim of this case-control study was to compare the efficacy of a multiple-dose GnRH antagonist protocol with that of the GnRH agonist long protocol with a view to oocyte and embryo quality, embryo development and IVF treatment outcome.</p> <p>Methods</p> <p>Between October 2001 and December 2008, 100 patients were stimulated with human menopausal gonadotrophin (HMG) and GnRH antagonist in their first treatment cycle for IVF or intracytoplasmic sperm injection (ICSI). One hundred combined GnRH agonist + HMG (long protocol) cycles were matched to the GnRH antagonist + HMG cycles by age, BMI, baseline FSH levels and by cause of infertility. We determined the number and quality of retrieved oocytes, the rate of early-cleavage embryos, the morphology and development of embryos, as well as clinical pregnancy rates. Statistical analysis was performed using Wilcoxon's matched pairs rank sum test and McNemar's chi-square test. P < 0.05 was considered statistically significant.</p> <p>Results</p> <p>The rate of cytoplasmic abnormalities in retrieved oocytes was significantly higher with the use of GnRH antagonist than in GnRH agonist cycles (62.1% vs. 49.9%; P < 0.01). We observed lower rate of zygotes showing normal pronuclear morphology (49.3% vs. 58.0%; P < 0.01), and higher cell-number of preembryos on day 2 after fertilization (4.28 vs. 4.03; P < 0.01) with the use of GnRH antagonist analogues. The rate of mature oocytes, rate of presence of multinucleated blastomers, amount of fragmentation in embryos and rate of early-cleaved embryos was similar in the two groups. Clinical pregnancy rate per embryo transfer was lower in the antagonist group than in the agonist group (30.8% vs. 40.4%) although this difference did not reach statistical significance (P = 0.17).</p> <p>Conclusion</p> <p>Antagonist seemed to influence favourably some parameters of early embryo development dynamics, while other morphological parameters seemed not to be altered according to GnRH analogue used for ovarian stimulation in IVF cycles.</p

Ion mobility spectrometry for the rapid analysis of over-the-counter drugs and beverages

In the pharmaceutical industry, there are increasing requirements for analytical methods in quality assessment for the production of drugs. In this investigation, ion mobility spectrometry (IMS) was used for the rapid qualitative separation and identification of active ingredients in generic over-the-counter drugs and food additives in beverages. The active ingredients determined in drugs were acetaminophen, aspartame, bisacodyl, caffeine, dextromethorphan, diphenhydramine, famotidine, glucosamine, guaifenesin, loratadine, niacin, phenylephrine, pyridoxine, thiamin, and tetrahydrozoline. Aspartame and caffeine were determined in beverages. Fourteen over-the-counter drugs and beverages were analyzed. Analysis times below 10 s were obtained for IMS, and reduced mobilities were reported for the first time for 12 compounds. A quadrupole mass spectrometer coupled to a mobility spectrometer was used to assure a correct peak assignation. The combination of fast analysis, low cost, and inexpensive maintenance of IMS instruments makes IMS an attractive technique for the qualitative determination of the active ingredients in over-the-counter drugs and food additives in manufacture quality control and cleaning verification for the drug and food industries

A Fiber-Optic Fluorescence Microscope Using a Consumer-Grade Digital Camera for In Vivo Cellular Imaging

BACKGROUND: Early detection is an essential component of cancer management. Unfortunately, visual examination can often be unreliable, and many settings lack the financial capital and infrastructure to operate PET, CT, and MRI systems. Moreover, the infrastructure and expense associated with surgical biopsy and microscopy are a challenge to establishing cancer screening/early detection programs in low-resource settings. Improvements in performance and declining costs have led to the availability of optoelectronic components, which can be used to develop low-cost diagnostic imaging devices for use at the point-of-care. Here, we demonstrate a fiber-optic fluorescence microscope using a consumer-grade camera for in vivo cellular imaging. METHODS: The fiber-optic fluorescence microscope includes an LED light, an objective lens, a fiber-optic bundle, and a consumer-grade digital camera. The system was used to image an oral cancer cell line labeled with 0.01% proflavine. A human tissue specimen was imaged following surgical resection, enabling dysplastic and cancerous regions to be evaluated. The oral mucosa of a healthy human subject was imaged in vivo, following topical application of 0.01% proflavine. FINDINGS: The fiber-optic microscope resolved individual nuclei in all specimens and tissues imaged. This capability allowed qualitative and quantitative differences between normal and precancerous or cancerous tissues to be identified. The optical efficiency of the system permitted imaging of the human oral mucosa in real time. CONCLUSION: Our results indicate this device as a useful tool to assist in the identification of early neoplastic changes in epithelial tissues. This portable, inexpensive unit may be particularly appropriate for use at the point-of-care in low-resource settings

PCR diagnostics and monitoring of adenoviral infections in hematopoietic stem cell transplantation recipients

After stem cell transplantation, human patients are prone to life-threatening opportunistic infections with a plethora of microorganisms. We report a retrospective study on 116 patients (98 children, 18 adults) who were transplanted in a pediatric bone marrow transplantation unit. Blood, urine and stool samples were collected and monitored for adenovirus (AdV) DNA using polymerase chain reaction (PCR) and real-time PCR (RT-PCR) on a regular basis. AdV DNA was detected in 52 (44.8%) patients, with mortality reaching 19% in this subgroup. Variables associated with adenovirus infection were transplantations from matched unrelated donors and older age of the recipient. An increased seasonal occurrence of adenoviral infections was observed in autumn and winter. Analysis of immune reconstitution showed a higher incidence of AdV infections during periods of low T-lymphocyte count. This study also showed a strong interaction between co-infections of AdV and BK polyomavirus in patients undergoing hematopoietic stem cell transplantations