A Circular RNA Expressed from the FAT3 Locus Regulates Neural Development.

Circular RNAs (circRNAs) are key regulators of cellular processes, are abundant in the nervous system, and have putative regulatory roles during neural differentiation. However, the knowledge about circRNA functions in brain development is limited. Here, using RNA-sequencing, we show that circRNA levels increased substantially over the course of differentiation of human embryonic stem cells into rostral and caudal neural progenitor cells (NPCs), including three of the most abundant circRNAs, ciRS-7, circRMST, and circFAT3. Knockdown of circFAT3 during early neural differentiation resulted in minor transcriptional alterations in bulk RNA analysis. However, single-cell transcriptomics of 30 and 90 days differentiated cerebral organoids deficient in circFAT3 showed a loss of telencephalic radial glial cells and mature cortical neurons, respectively. Furthermore, non-telencephalic NPCs in cerebral organoids showed changes in the expression of genes involved in neural differentiation and migration, including FAT4, ERBB4, UNC5C, and DCC. In vivo depletion of circFat3 in mouse prefrontal cortex using in utero electroporation led to alterations in the positioning of the electroporated cells within the neocortex. Overall, these findings suggest a conserved role for circFAT3 in neural development involving the formation of anterior cell types, neuronal differentiation, or migration.

Terminal uridyltransferase 7 regulates TLR4-triggered inflammation by controlling Regnase-1 mRNA uridylation and degradation.

Different levels of regulatory mechanisms, including posttranscriptional regulation, are needed to elaborately regulate inflammatory responses to prevent harmful effects. Terminal uridyltransferase 7 (TUT7) controls RNA stability by adding uridines to its 3' ends, but its function in innate immune response remains obscure. Here we reveal that TLR4 activation induces TUT7, which in turn selectively regulates the production of a subset of cytokines, including Interleukin 6 (IL-6). TUT7 regulates IL-6 expression by controlling ribonuclease Regnase-1 mRNA (encoded by Zc3h12a gene) stability. Mechanistically, TLR4 activation causes TUT7 to bind directly to the stem-loop structure on Zc3h12a 3'-UTR, thereby promotes Zc3h12a uridylation and degradation. Zc3h12a from LPS-treated TUT7-sufficient macrophages possesses increased oligo-uridylated ends with shorter poly(A) tails, whereas oligo-uridylated Zc3h12a is significantly reduced in Tut7-/- cells after TLR4 activation. Together, our findings reveal the functional role of TUT7 in sculpting TLR4-driven responses by modulating mRNA stability of a selected set of inflammatory mediators.

Trends in the use of antimuscarinics and alpha-adrenergic blockers in women with lower urinary tract symptoms in Taiwan: A nationwide, population-based study, 2007-2012.

BACKGROUND: We aim to examine the trend in the use of antimuscarinics and off-label alpha-adrenergic blockers for treatment of lower urinary tract symptoms (LUTS) in a Taiwanese Women Cohort between 2007 and 2012. METHODS: This population-based National Health Insurance Research Database (NHIRD) was used to examine the trends in the use of antimuscarinics or off-label alpha-adrenergic blockers in Taiwan. A sample of 1,000,000 individuals randomly drawn from the whole population of 23 million individuals who were registered in the NHI in 2005. From 2007 through 2012, women aged over 18 years whose claim record contained prescriptions of either of the two drugs for treatment of any of the LUTS-related diagnoses were identified and analyzed. The annual usage of the two drug classes were calculated by defined daily dose (DDD). RESULTS: From 2007-2012, there was a 0.80 fold (69676.8 to 125104.3) increase in DDD of antimuscarinics in our cohort. The overall healthcare seeking prevalence of LUTS was 7.33% in 2007 and 12.38% in 2012, in a rising trend. The prevalence of antimuscarinics-treated LUTS in our cohort increased from 2.53 in 2007 to 3.41 per 1000 women in 2012. The prevalence of LUTS treated by antimuscarinics increased especially for those older than 60 years during the study period. CONCLUSIONS: This 6-year observational study provided the epidemiologic information of clinically significant LUTS of Asian female population. Moreover, there was a rising trend in the use of antimuscarinics and off-label alpha-adrenergic blockers in the population-based cohort.

Isolating, Sequencing and Analyzing Extracellular MicroRNAs from Human Mesenchymal Stem Cells.

Extracellular and circulating RNAs (exRNA) are produced by many cell types of the body and exist in numerous bodily fluids such as saliva, plasma, serum, milk and urine. One subset of these RNAs are the posttranscriptional regulators - microRNAs (miRNAs). To delineate the miRNAs produced by specific cell types, in vitro culture systems can be used to harvest and profile exRNAs derived from one subset of cells. The secreted factors of mesenchymal stem cells are implicated in alleviating numerous diseases and is used as the in vitro model system here. This paper describes the process of collection, purification of small RNA and library generation to sequence extracellular miRNAs. ExRNAs from culture media differ from cellular RNA by being low RNA input samples, which calls for optimized procedures. This protocol provides a comprehensive guide to small exRNA sequencing from culture media, showing quality control checkpoints at each step during exRNA purification and sequencing.

Factors Associated with Bothersome Lower Urinary Tract Symptoms in Middle-Aged Men Receiving Health Checkup.

The prospective study is to investigate the associations between serum testosterone levels and LUTS among middle-aged men ≥40 years receiving health check-up. Lower urinary tract symptoms were evaluated by the self-administered International Prostate Symptom Score questionnaire. Serum prostate specific antigen and total testosterone level were checked in all subjects. A total of 1752 men were enrolled into the study. The mean age was 55.6 ± 9.7 years. All study subjects were stratified into low, medium and high testosterone levels by two cut-off, 3.0 and 4.11 ng/mL. We found that testosterone levels were significantly associated with metabolic syndrome and body fat components. Compared to those with low testosterone levels, subjects with high and medium testosterone had a significantly higher IPSS (5.84 ± 5.55 vs 6.71 ± 5.68 and 6.34 ± 5.66, p = 0.032) and storage score (2.76 ± 2.29 vs 3.20 ± 2.49 and 2.90 ± 2.49; p = 0.009), and a more moderate/severe LUTS (IPSS ≧ 8) (26.5% vs 35.7% and 29.9%; p = 0.002). Multivariate analyses showed that high vs low testosterone levels (OR, 1.76; 95% CI, 1.26-2.45) and prostate volume ≧25 vs <25 mL (OR, 1.38; 95% CI, 1.04-1.82) significantly associated with the presence of moderate/severe LUTS. Pearson correlation analyses showed significantly positive correlations between testosterone level and IPSS in whole study sample (Pearson correlation coefficient, 0.066; p < 0.01) and in the subgroup of moderate/severe LUTS (Pearson correlation coefficient, 0.038; p < 0.05). In conclusion, high testosterone and prostate volume adversely impacted LUTS in our target population.

Caveolae-mediated mesenchymal stem cell labelling by PSS-coated PLGA PFOB nano-contrast agent for MRI.

Rationale: Non-invasive tracking of transplanted cells is critical in evaluating delivery, migration and prognosis of cell therapies. Methods: We formulated a nano-contrast agent consisting of a perfluorooctylbromide (PFOB) core within a shell of poly (lactic-co-glycolic acid) (PLGA) followed by a coat of polystyrene sulfonate (PSS) for 19F MRI. The nano-contrast agent (PSS-NP) was characterised by DLS and the uptake efficiency of the nano-contrast agent (PSS-NP) was tested using flow cytometry, in vitro MRI and confocal microscopy. In vitro and in vivo assays of labelled cells were tested for their ability to provide an MRI signal while retaining their osteoblastic differentiation capabilities. Results: PSS-NPs were internalised via caveolae-mediated endocytosis in mesenchymal stromal/stem cells without affecting cell proliferation and differentiation in osteoblasts, both in vitro and in vivo. Furthermore, labelled cells were monitored by 19F MRI for up to 2 months after transplantation in mice. In particular, PSS-NP-labelled cells can be used to monitor the enhanced immune rejection of grafted human cells in normal BALB/c mice compared to immune-compromised NOD/SCID mice. One week after transplantation, 40% of the 19F MRI signal was lost in normal mice, whereas only 10% was lost in immune-compromised mice. Conclusion: Overall, these results show that PSS-NPs can label MSCs effectively, and be employed in vivo as a novel nano-contrast agent for non-invasive cell tracking using clinically relevant 19F MRI techniques.

Global MicroRNA Profiling in Human Bone Marrow Skeletal-Stromal or Mesenchymal-Stem Cells Identified Candidates for Bone Regeneration.

Bone remodeling and regeneration are highly regulated multistep processes involving posttranscriptional regulation by microRNAs (miRNAs). Here, we performed a global profiling of differentially expressed miRNAs in bone-marrow-derived skeletal cells (BMSCs; also known as stromal or mesenchymal stem cells) during in vitro osteoblast differentiation. We functionally validated the regulatory effects of several miRNAs on osteoblast differentiation and identified 15 miRNAs, most significantly miR-222 and miR-423, as regulators of osteoblastogenesis. In addition, we tested the possible targeting of miRNAs for enhancing bone tissue regeneration. Scaffolds functionalized with miRNA nano-carriers enhanced osteoblastogenesis in 3D culture and retained this ability at least 2 weeks after storage. Additionally, anti-miR-222 enhanced in vivo ectopic bone formation through targeting the cell-cycle inhibitor CDKN1B (cyclin-dependent kinase inhibitor 1B). A number of additional miRNAs exerted additive osteoinductive effects on BMSC differentiation, suggesting that pools of miRNAs delivered locally from an implanted scaffold can provide a promising approach for enhanced bone regeneration.

Intimacy in Phone Conversations: Anxiety Reduction for Danish Seniors with Hugvie.

There is a lack of physical contact in current telecommunications such as text messaging and Internet access. To challenge the limitation and re-embody telecommunication, researchers have attempted to introduce tactile stimulation to media and developed huggable devices. Previous experiments in Japan showed that a huggable communication technology, i.e., Hugvie decreased stress level of its female users. In the present experiment in Denmark, we aim to investigate (i) whether Hugvie can decrease stress cross-culturally, i.e., Japanese vs. Danish participants (ii), investigate whether gender plays a role in this psychological effect (stress reduction) and (iii) if there is a preference of this type of communication technology (Hugvie vs. a regular telephone). Twenty-nine healthy elderly participated (15 female and 14 male, M = 64.52 years, SD = 5.67) in Jutland, Denmark. The participants filled out questionnaires including State-Trait Anxiety Inventory, NEO Five Factor Inventory (NEO-FFI), and Becks Depression Inventory, had a 15 min conversation via phone or Hugvie and were interviewed afterward. They spoke with an unknown person of opposite gender during the conversation; the same two conversation partners were used during the experiment and the Phone and Hugvie groups were equally balanced. There was no baseline difference between the Hugvie and Phone groups on age or anxiety or depression scores. In the Hugvie group, there was a statistically significant reduction on state anxiety after meeting Hugvie (p = 0.013). The change in state anxiety for the Hugvie group was positively correlated with openness (r = 0.532, p = 0.041) as measured by the NEO-FFI. This indicates that openness to experiences may increase the chances of having an anxiety reduction from being with Hugvie. Based on the results, we see that personality may affect the participants' engagement and benefits from Hugvie. We discuss the implications of the results and further elaborations.

Peracylated Glucosyl Kaempferols from Pasania dodonfifolia Leaf.

Phytochemical investigation of the ethanolic extract of Pasania dodoniifolia leaf led to the isolation of four kaempferol 3-0-peracylated glucosides (1-4), together with four flavonoid glucosides (5-8), epicatechin (9), and (7S, 7'S, 8R, 8'R)-icariol A2 (10). Of these, kaempferol-3-O-(3",4"-di-O-acetyl-2"-O-(Z)-p- coumaroyl)-6"-O-(E)-p-coumaroyl)-beta-glucopyranoside (3) and 3-O-(3",4"-di-O-acetyl-2",6"-di-O-(Z)-p-coumaroyl)-beta-glucopyranoside (4) are new and their structures were elucidated by 2D NMR spectroscopic analyses and MS data.

Acylated glucosylflavones as α-glucosidase inhibitors from Tinospora crispa leaf.

The ethanolic extract of Tinospora crispa leaf had shown inhibitory activity toward α-glucosidase. Bioassay guided fractionation and separation of this extract led to the isolation of 17 flavonoids. Among them, four acylated glycosylflavonoids (6, 8, 9, 15) are new compounds. Their structures were elucidated on the basis of spectroscopic analysis. Among the isolated compounds, isovitexin 2″-(E)-p-coumarate (8) showed the best activity against α-glucosidase with an IC50 value of 4.3±1.4µM. However, isoorientin 2″-(E)-p-coumarate (7), the 3'-hydroxylated 8, is much less active (IC50 35.7µM). Such significant difference was rationalized by CAD study on α-glucosidase.

The beneficial effect of Batroxobin on blood loss reduction in spinal fusion surgery: a prospective, randomized, double-blind, placebo-controlled study.

OBJECTIVE: Our objective was to evaluate the efficacy and safety of Batroxobin on blood loss during spinal operations. METHODS: After obtaining approval from the ethics committee at the hospital along with informed written consent, we performed a double-blind, randomized, placebo-controlled study with 100 patients who were randomized equally into 2 groups (Batroxobin and placebo). Patients received either 2 ku IV 15 min before surgery and followed 1 ku IM of Batroxobin following surgery, or an equivalent volume of placebo (normal saline). Cost of Batroxobin treatment is amounted to 84.75 euros. The primary outcomes were intraoperative, 24 h postoperative, and total perioperative blood loss. Secondary outcomes were hemoglobin (Hb), red blood cell count (RBC), the volume of blood/fluid transfusion intraoperatively, and 24 h postoperatively. Safety evaluation parameters were the incidence of venous thrombosis in the lower extremities, active partial thromboplastin time, prothrombin time, thrombin time, and fibrinogen. The data were analyzed using the Statistical Package for the Social Science Version 12.0. The results were presented as mean ± SEM. The Mann-Whitney test and Independent Student t test, when appropriate, were used to compare the 2 groups, and differences were considered significant if the P value was <0.05. RESULTS: 88 patients were included in the analysis while 12 patients were withdrawn from the study due to extended surgical duration, change of surgical procedure, or after the patients' request. The total perioperative blood loss was approximately 31% lower in patients given Batroxobin versus placebo (700.5 ± 45.81 vs 485.7 ± 30.01 mL, P = 0.001). The Batroxobin group had significantly less intraoperative blood loss (326.1 ± 24.16) compared to the placebo group (556.0 ± 43.58), but there was no difference in the amount of blood/fluid transfused, postoperatively Hb, or RBC between the two groups. After the operation, coagulation parameters were not significantly different between the 2 groups at the days 1 or 3 postoperatively. No adverse events related to the use of Batroxobin were recorded. There were no cases of superficial wound infection. None of the subjects died during the study. CONCLUSIONS: In this study, prophylactic use of Batroxobin provided an effective and cheap method for reducing blood loss without coagulopathy during or after operations. The use of Batroxobin for patients undergoing one-level PLIF surgery safely and effectively reduced the total amount of perioperative blood loss.

Episodic retrieval and decaying inhibition in the competitor-rule suppression phenomenon.

The Competitor Rule Suppression (CRS) effect is the performance impairment observed in task switching when the currently relevant task rule is the same rule that had generated a response conflict in the preceding trial. This effect could reflect (a) episodic tagging, in which a competitor rule is retrieved with relative difficulty in subsequent trials or (b) residual active inhibition of the competing rule. In order to help distinguishing between the two accounts, the authors manipulated the Response-Cue Interval (RCI), which may influence both processes. CRS increased with increasing temporal distinctiveness between the previous and current episode (operationalized by the ratio of the current RCI to the previous RCI, RCI/pRCI), thus supporting episodic tagging. CRS additionally decreased numerically with increasing RCI even when the RCI/pRCI ratio was fixed, thereby providing suggestive support for the decay account.

"Smart inhibition": electrophysiological evidence for the suppression of conflict-generating task rules during task switching.

A major challenge for task switching is maintaining a balance between high task readiness and effectively ignoring irrelevant task rules. This calls for finely tuned inhibition that targets only the source of interference without adversely influencing other task-related representations. The authors show that irrelevant task rules generating response conflict are inhibited, causing their inefficient execution on the next trial (indicating the presence of competitor rule suppression[CRS];Meiran, Hsieh, & Dimov, Journal of Experimental Psychology: Learning, Memory and Cognition, 36, 992-1002, 2010). To determine whether CRS influences task rules, rather than target stimuli or responses, the authors focused on the processing of the task cue before the target stimulus was presented and before the response could be chosen. As was predicted, CRS was found in the event-related potentials in two time windows during task cue processing. It was also found in three time windows after target presentation. Source localization analyses suggest the involvement of the right dorsal prefrontal cortex in all five time windows.

Left visual-field advantage in the dual-stream RSVP task and reading-direction: a study in three nations.

In the dual-stream Rapid Serial Visual Presentation task, a stream of stimuli containing two target stimuli is rapidly presented left and right. In previous studies, the second target was better identified in the left than in the right hemifield. In all those studies, alphanumeric stimuli were used both as targets and distracters. We examined to what extent this left visual-field advantage is dependent on reading-direction. The task was performed by Germans (with Latin characters), Israelis (with Latin and Hebrew characters) and Taiwanese (with Latin and Chinese characters). If caused by overlearnt associative links between Latin characters and left-to-right reading, the prominent left visual-field bias should be reversed in Hebrew and disappear in Chinese. Furthermore, if caused by direction of reading in the participant's native language, the left visual-field advantage in Latin conditions should be larger in Germans than in Israelis and Taiwanese. A left visual-field advantage was always observed, though slightly smaller in Hebrew and in Chinese, and there was no difference in the Latin conditions between the three nations. Therefore, it seems that the left visual-field advantage in speeded target identification is not primarily caused by the left-to-right reading-direction, but may be a combined effect resulting from the asymmetric organization of general mechanisms of visual processing and from stimulus-induced preferences.