DICER1 -Altered Extraovarian Moderately Differentiated Sertoli-Leydig Cell Tumor: Report of a Rare Case.

We report an unusual case of a pelvic extraovarian moderately differentiated Sertoli-Leydig cell tumor arising in a 4-yr-old female. The tumor contained a DICER1 pathogenic variant which was absent in the germline ruling out DICER1 syndrome. In reporting this case, we discuss the differential diagnosis and possible histogenesis and review reported cases of extraovarian Sertoli-Leydig cell tumor.

On the Quality, Statistical Efficiency, and Safety of Simultaneously Recorded Multiband fMRI/EEG.

The recent development of multiband functional magnetic resonance imaging (MB-fMRI) allows for the reduction of sampling period by simultaneously exciting multiple slices-the number of which is referred to as the multiband factor. Simultaneously recorded electroencephalography (EEG)/MB-fMRI has yet to be validated for data quality against conventional single band (SB)-fMRI. Pilot scans were conducted on phantoms twice and on a healthy volunteer to ensure no heating effects. In the main study, two thermometer probes were attached to 16 healthy individuals (ages 20-39, 9 females) whilst they completed two sets of 16-min resting-state and two sets of 9-min n-back task scans-each set consisting of one MB4 and one SB pulse sequence. No heating effects were reported and thermometer data showed mean increases of < 1.0 °C. Minimal differences between the two scan types were found in EEG channel variance and spectra. Expected decreases in MB4-fMRI tSNR were observed. In n-back task scans, little to no differences were detected in both EEG source analyses and fMRI local analyses for mixed effects. Resting-state posterior cingulate cortex seed-based analyses of the default mode network along with EEG-informed fMRI analysis of the occipital alpha anticorrelation effect showed improved statistical and spatial sensitivity at lower scan durations. Using EEG/MB4-fMRI for n-back tasks provided no statistical advantages nor disadvantages. However, for studying the resting-state, MB4-fMRI potentially allows for reduced scanning durations for equivalent statistical significance to be obtained or alternatively, larger effect sizes for the same scanning duration. As such, simultaneous EEG/MB4-fMRI is a viable alternative to EEG/SB-fMRI.

Genome-Wide Transcriptome Profiling of the Neoplastic Giant Cell Tumor of Bone Stromal Cells by RNA Sequencing.

Giant cell tumor of bone (GCTB) is the most common non-malignant primary bone tumor reported in Hong Kong. Failure of treatment in advanced GCTB with aggressive local recurrence remains a clinical challenge. In order to reveal the molecular mechanism underlying the pathogenesis of this tumor, we aimed to examine the transcriptome profiling of the neoplastic stromal cells of GCTB in this study. RNA-sequencing was performed on three GCTB stromal cell samples and one bone marrow-derived MSC sample and 174 differentially expressed genes (DEGs) were identified between these two cell types. The top five up-regulated genes are SPP1, F3, TSPAN12, MMP13, and LGALS3BP and further validated by qPCR and Western Blotting. Knockdown of SPP1 was found to induce RUNX2 and OPG expression in GCTB stromal cells but not the MSCs. Ingenuity pathway analysis (IPA) of the 174 DEGs revealed significant alternations in 23 pathways; variant calling analysis revealed 1915 somatic variants of 384 genes with high or moderate impacts. Interestingly, four canonical pathways were found overlapping in both analyses; from which VEGFA, CSF1, PLAUR, and F3 genes with somatic mutation were found up-regulated in GCTB stromal cells. The STRING diagram showed two main clusters of the DEGs; one cluster of histone genes that are down-regulated in GCTB samples and another related to osteoblast differentiation, angiogenesis, cell cycle progression, and tumor growth. The DEGs and somatic mutations found in our study warrant further investigation and validation, nevertheless, our study add new insights in the search for new therapeutic targets in treating GCTB. J. Cell. Biochem. 118: 1349-1360, 2017. © 2016 Wiley Periodicals, Inc.

Profiling of the silica-induced molecular events in lung epithelial cells using the RNA-Seq approach.

Silicosis is a prolonged, irreversible and incurable occupational disease, and there is a significant number of newly diagnosed cases every year in Hong Kong. Due to the long latency of the disease, the diagnosis can be missed until detailed clinical examination at a later stage. For a better control of this deadly disease, detailing the pro-inflammatory and fibrotic events in the macrophage would be instrumental in understanding the pathogenesis of the disease and essential for the significant biomarkers discovery. In this in vitro study, human cell line model A549 lung epithelial cells were used. The immediate molecular events underneath the activation of quartz silica polymorphs were followed in a time course of 0, 0.5, 2, 8, 16 and 24 h. The transcriptome library was prepared and subjected to RNA-Seq analysis. Data analysis was performed by pathway analysis tools and verified by real-time PCR. The results showed that triggered genes were mainly found in the immune response and inflammatory pathways. An interesting finding was the association of the DNA-binding protein inhibitor (ID) family in the silica exposure to lung cells. The linkage of ID1, ID2 and ID3 to cancer may rationalize themselves to be the markers indicating an early response of silicosis. However, further studies are required to consolidate the roles of these genes in silicosis. Copyright © 2017 John Wiley & Sons, Ltd.

A Randomized Controlled Trial of Intravenous Scopolamine Versus Active-Placebo Glycopyrrolate in Patients With Major Depressive Disorder.

Objective: To investigate scopolamine's rapid-acting antidepressant effects using an active placebo comparator. Most prior intravenous scopolamine studies reduced depressive symptomatologies compared to saline placebo infusions within 3 days. However, the confounding effect of placebo is unknown given that only saline placebo has been used in prior studies.Methods: In this trial, 40 patients with major depressive disorder were randomized to receive single intravenous doses of either scopolamine hydrobromide (4-6 µg/kg) or glycopyrronium bromide (4 µg/kg) between August 2019 and April 2021 in Auckland, New Zealand. Glycopyrronium was chosen as the active placebo due to its similar antimuscarinic properties to scopolamine but inability to cross the blood-brain barrier. The primary mood outcome measure was the Montgomery-Åsberg Depression Rating Scale (MADRS) administered pre-infusion and 1, 3, 7, 14, 28, and 42 days post-infusion.Results: Per protocol, this trial was abandoned for futility at n = 40. While scopolamine reduced MADRS scores by 12.6 (± 8.7 SD) points at day 3, glycopyrronium showed similar reductions (11.2 ± 9.6 SD). Frequentist linear mixed models showed no antidepressant effects of scopolamine versus placebo (d = 0.17), and Bayesian mixed effect models showed moderate evidence in favor of the null hypothesis at day 3 (Bayes factor = 0.32). Participants remained well-blinded to drug allocation, with 50% of participants correctly guessing their allocation.Conclusions: The observed MADRS improvement was larger than in prior studies, but no antidepressant effects were observed. This study using an active placebo confirms recent studies demonstrating the lack of antidepressant efficacy of scopolamine.Trial Registration: Australian New Zealand Clinical Trials Registry identifier: ACTRN12619000569101.

Extent of resection affects prognosis for patients with glioblastoma in non-eloquent regions.

Glioblastoma (GBM) is a malignant cerebral neoplasm carrying poor prognosis. The importance of extent of resection (EoR) in GBM patient outcomes has been argued in the literature. Previous studies included tumors in eloquent regions of the brain. This confounds the role of EoR by including patients with intrinsically worse outcomes but will be over-represented in the reduced EoR category. In a homogenous group of patients in whom GTR was considered achievable, we investigated the effect of increasing EoR on survival. A retrospective review of 51 patients was undertaken. Quantitative, volumetric analysis of pre-operative and post-operative magnetic resonance image was compared with corresponding clinical details. The primary outcome measured was post-operative overall survival. Median overall survival was 18.3 months for GTR patients compared to 11.6 months for non-GTR (p = 0.025). Median pre-operative contrast-enhancing tumor volume for GTR patients was 54.7 cm3 and 24.9 cm3 for non-GTR. Post-operative median residual tumor volume was 1.1 cm3 in the non-GTR cohort. In multivariate analyses, GTR (HR [95% CI] = 0.973 [0.954-0.994], p = 0.00559) and increasing EoR (HR [95% CI] = 0.964 [0.944-0.985], p = 0.000665) remained predictors of survival. Centile dichotomization of EoR revealed 74% (HR [95% CI] = 0.351 [0.128-0.958], p = 0.0409) as the lowest threshold conferring statistically significant survival benefit. Where technically feasible, both GTR and EoR remained as independent prognostic factors for survival. GTR remains the gold standard for surgical treatment of GBM in patients, 74% being the minimum EoR required to confer survival benefit.

A randomised, double-blind, active placebo-controlled, parallel groups, dose-response study of scopolamine hydrobromide (4-6 µg/kg) in patients with major depressive disorder.

BACKGROUND: Depressive disorders are a leading cause of disability, but current behavioural and pharmacological therapies have a slow onset of response, typically taking several weeks before achieving efficacy. Prior studies using triplicate intravenous scopolamine infusions have been shown to reduce depressive symptomologies within days compared to saline placebo infusions. However, several parameters of scopolamine's potential antidepressant effect remain unknown, such as its dose-response profile and its washout period. There is also the question as to whether the previously reported antidepressant responses were confounded by unblinding effects due to the lack of an active placebo control. Glycopyrronium bromide was selected as placebo for this trial given it has similar antimuscarinic properties to scopolamine hydrobromide but an inability to cross the blood-brain barrier, thereby hypothetically mimicking only the peripheral effects of scopolamine. METHODS/DESIGN: A parallel group trial of single intravenous scopolamine infusions at three doses (4, 5, and 6 µg/kg) along with one glycopyrronium bromide 4 µg/kg group will be administered to 40 participants with major depressive disorder in a 1:1:1:2 ratio, respectively. The primary outcome measure will be the Montgomery-Åsberg Depression Rating Scale (MADRS) administered at baseline, 4 hours, 1 day, 3 days, 1 week, 2 weeks, 4 weeks, and 6 weeks post-infusion to determine antidepressant efficacy. As a secondary measure, the Quick Inventory of Depressive Symptomatology will be administered alongside the MADRS to further track potential antidepressant responses. Other secondary measures include electroencephalography, blood samples, and Bowdle visual acuity scales recorded at baseline, 5, 10, 15, 20, 30, 60, 120, and 240 min post-infusion to determine the pharmacokinetic-pharmacodynamic profile of scopolamine in depressed participants. DISCUSSION: This trial contributes to the literature surrounding the efficacy of scopolamine as an antidepressant. Determining the dose-response profile and washout period of scopolamine's antidepressant effect will also provide important information for designing and conducting crossover trials. The use of an active placebo is important to reduce potentially confounding expectancy effects. TRIAL REGISTRATION: The trial was registered in the Australian New Zealand Clinical Trials Registry (registration number ACTRN12619000569101). Registered on 11 April 2019.

RNA-Seq revealed ATF3-regulated inflammation induced by silica.

BACKGROUND: Millions of workers are exposed to dust containing silica. Chronic and over-exposure to silica will lead to silicosis, which is an irreversible and sometimes fatal lung disease. The disordered physiological processes of silicosis consist of accumulation of silica particles in the alveoli of the lung. Then, the ingestion of the silica particles by macrophages was followed by an inflammatory response. Up till now, the chest radiographs remain the key tool in diagnosing and assessing the extent of silicosis. However, concerns exist regarding the sensitivity and specificity of the technique. Therefore, there is still a need to develop a biomarker for silicosis for early detection of silicosis. METHOD: In this study, RNA-Seq was applied to detect the gene expression changes when silica was exposed to macrophages at different time intervals. RNA-Seq provides a broader dynamic range, increased specificity and sensitivity, and easier detection of rare and low-abundance transcripts. Bioinformatics tools such as the Database for Annotation, Visualization and Integrated Discovery (DAVID) and Gene Functional Classification Tool and Search Tool for the Retrieval of Interacting Genes (STRING) were applied for data analysis. Quantitative PCR was used to validate the results. RESULTS: Our results showed that regulation of transcription factors was the dominant activated pathway in early exposure of silica to macrophages, followed by inflammatory responses which were the main mechanisms in silicosis. One of the findings was the upregulation of activating transcription factor 3 (ATF3) during silica exposure. When ATF3 expression was inhibited by siRNA, the production of cytokines IL-1ß, IL-6 and TNF was further increased. CONCLUSION: This indicated that ATF3 may be a potential early diagnostic biomarker for silicosis and ATF3 acts as a repressor in inflammatory responses induced by silica.

Medicinal plants used by Tanzanian traditional healers in the management of Candida infections.

An ethnomedical survey in Coast, Dar es Salaam, Morogoro and Tanga regions of Tanzania has resulted in the identification of 36 plant species belonging to 21 plant families that are used traditionally for the treatment of Candida infections. Twenty-one plants constituting 58.3% of all collected plants are used to treat of oral candidiasis (Utando) one of the important signs of HIV/AIDS. The knowledge of traditional healers for the treatment of Candida infections has been highly supported by the literature in that 13 (36.1%) out of the 36 plants identified have been proven to be active against Candida albicans and/or other species of Candida. Also, some of the plants were reported to be active against other species of fungi including Cryptococcus neoformans, one of the important pathogenic fungi in HIV/AIDS. It can be seen that ethnomedical information from traditional healers provides a solid lead towards development of new drugs than random screening. The task that remains is to screen extracts prepared from these plants and perform a bioassay-guided fractionation of the active extracts so as to isolate the active compounds from these plants.

Larvicidal and mosquitocidal extracts, a coumarin, isoflavonoids and pterocarpans from Neorautanenia mitis.

The present study aimed to evaluate extracts and compounds from tubers of Neorautanenia mitis against the malaria- and filariasis-transmitting mosquitoes, Anopheles gambiae and Culex quinquefaciatus, respectively. The extracts exhibited activity against larvae of A. gambiae and C. quinquefaciatus mosquitoes, and were also active against adult A. gambiae mosquitoes. The active extracts yielded the coumarin derivative pachyrrhizine, the isoflavonoids neotenone and neorautanone, and the pterocarpans neoduline, nepseudin and 4-methoxyneoduline as the active constituents. The activity of the crude extracts was at about the same magnitude as that of the constituent natural products, the latter's efficacy being almost at the same level for all the isolated compounds. The mosquitocidal activities of the pure compounds were comparable to those of the standard mosquitocides deltamethrin and alphacypermethrin. These findings corroborate traditional insecticidal application of N. mitis and the results can be extended for the control of mosquitoes especially at breeding sites.

Isolation of a stilbene glycoside and other constituents of Terminalia sericeae.

The ethanol extract of the root bark of Terminalia sericea yielded an unreported stilbene glycoside, 3'5'-dihydroxy-4-(2-hydroxy-ethoxy) resveratrol-3-O-beta-rutinoside (1) together with known compounds resveratrol-3-beta-rutinoside glycoside (2), 3',4,5'-Trihydroxystilbene (resveratrol) (3), triterpenoic acid arjungenin and a mixture of beta-sitosterol and stigmasterol. Structure determination of the isolated compounds was achieved on the basis of spectroscopic measurements.

In vitro antimicrobial assay of plants used in traditional medicine in Bukoba Rural district, Tanzania.

Plants used in traditional medicine in Bukoba Rural district in Tanzania were evaluated for their in vitro antimicrobial activities. Plant materials from eight plant species (Harungana madagascariensis (Lam) Poir., Jatropha curcas L., Lantana trifolia L., Plectranthus barbatus Andr., Pseudospondias microcarpa Engl., Psorospermum febrifugum Spach, Teclea nobilis Del. and Vernonia adoensis [Warp.] SL) were collected based on ethnomedical information provided by traditional herbal practitioners. Results of the study indicate that extracts from the eight plant species were active against at least one or more of the test organisms (Bacillus subtilis, Staphylococcus aureus [gram positive], Escherichia coli, Pseudomonas aeruginosa [gram negative] and Candida albicans [Yeast]). A profile of secondary metabolites (alkaloids, terpenoids, triterpenes, phenolics, tannins, flavonoids, anthraquinones, flavonols/flavones and /or chalcones, sterols and saponins) was obtained for three plant species (Jatropha curcas L., Plectranthus barbatus Andr., and Pseudospondias microcarpa Engl.). The paper discusses the probable therapeutic basis of these traditional plants based on their secondary metabolite profiles and for the first time draws research attention to Bukoba Rural district as a source for plants with potential pharmaceutical applications.

Action of adenosine triphosphate on human skin and gingival fibroblasts. Protective action of fibronectin.

If cultured in media supplemented with adenosine triphosphate (ATP), EDTA, trypsin, thrombin, or incubated at 0-15 degrees C, human skin fibroblasts (HSF) and human gingival fibroblasts (HGF) change from long attached elliptical to round floating cell cultures. Also, if treated with ATP, EDTA, trypsin, thrombin, or incubated at 0-15 degrees C, the attached HFS or HGF monolayers detach from plastic substratum and form floating round cells that progressively aggregate together and die. The described experiments examined the role of cellular and extracellular ATP on HSF and HGF attachment. These two types of fibroblasts differed in their cellular ATP levels and their response to metabolic inhibitors. ATP causes destruction of microtubules as monitored by colcemid uptake and cellular detachment. Fibronectin protects both HSF and HGF from the effects of extracellular ATP.

Fluoxetine-induced memory impairment in four family members.

OBJECTIVE: Fluoxetine, a selective serotonin reuptake inhibitor antidepressant agent, has been implicated in learning and memory. Here, we report four cases from the same family of fluoxetine-related memory-impairment. RESULTS: Memory-impairment resulted after fluoxetine treatment and disappeared after changing to another selective serotonin reuptake inhibitor. CONCLUSIONS: There may be a relationship between fluoxetine-related memory impairment and genetic factors, and this side-effect appears to be specific to fluoxetine treatment. Possible mechanisms underlying this effect may be the drug's influence on the central serotonergic system or brain-derived neurotrophic factor.