Tamoxifen, an estrogen receptor (ER) antagonist, is the mainstay treatment of breast cancer and the development of resistance represents a major obstacle for a cure. Although long non-coding RNAs ...such as HOTAIR have been implicated in breast tumorigenesis, their roles in chemotherapy resistance remain largely unknown. In this study, we report that HOTAIR (HOX antisense intergenic RNA) is upregulated in tamoxifen-resistant breast cancer tissues compared to their primary counterparts. Mechanistically, HOTAIR is a direct target of ER-mediated transcriptional repression and is thus restored upon the blockade of ER signaling, either by hormone deprivation or by tamoxifen treatment. Interestingly, this elevated HOTAIR increases ER protein level and thus enhances ER occupancy on the chromatin and potentiates its downstream gene regulation. HOTAIR overexpression is sufficient to activate the ER transcriptional program even under hormone-deprived conditions. Functionally, we found that HOTAIR overexpression increases breast cancer cell proliferation, whereas its depletion significantly impairs cell survival and abolishes tamoxifen-resistant cell growth. In conclusion, the long non-coding RNA HOTAIR is directly repressed by ER and its upregulation promotes ligand-independent ER activities and contributes to tamoxifen resistance.
We have previously shown that miR-486-5p is one of the most downregulated micro RNAs in lung cancer. The objective of the study was to investigate the role of miR-486-5p in the progression and ...metastasis of non-small-cell lung cancer (NSCLC). We evaluated miR-486-5p expression status on 76 frozen and 33 formalin-fixed paraffin-embedded tissues of NSCLC by quantitative reverse transcriptase PCR to determine its clinicopathologic significance. We then performed function analysis of miR-486-5p to determine its potential roles on cancer cell migration and invasion in vitro and metastasis in vivo. We also investigated the target genes of miR-486-5p in lung tumorigenesis. miR-486-5p expression level was significantly lower in lung tumors compared with their corresponding normal tissues (P<0.0001), and associated with stage (P=0.0001) and lymph node metastasis of NSCLC (P=0.0019). Forced expression of miR-486-5p inhibited NSCLC cell migration and invasion in vitro and metastasis in mice by inhibiting cell proliferation. Furthermore, ectopic expression of miR-486-5p in cancer cells reduced ARHGAP5 expression level, whereas miR-486-5p silencing increased its expression. Luciferase assay demonstrated that miR-486-5p could directly bind to the 3'-untranslated region of ARHGAP5. The expression level of miR-486-5p was inversely correlated with that of ARHGAP5 in lung tumor tissues (P=0.0156). Reduced expression of ARHGAP5 considerably inhibited lung cancer cell migration and invasion, resembling that of miR-486-5p overexpression. miR-486-5p may act as a tumor-suppressor contributing to the progression and metastasis of NSCLC by targeting ARHGAP5. miR-486-5p would provide potential diagnostic and therapeutic targets for the disease.
While oral antidepressants reach efficacy after weeks, single-dose intravenous (i.v.) ketamine has rapid, yet time-limited antidepressant effects. We aimed to determine the efficacy and safety of ...single-dose i.v. ketamine augmentation of escitalopram in major depressive disorder (MDD).
Thirty outpatients with severe MDD (17-item Hamilton Rating Scale for Depression total score ⩾ 24) were randomized to 4 weeks double-blind treatment with escitalopram 10 mg/day+single-dose i.v. ketamine (0.5 mg/kg over 40 min) or escitalopram 10 mg/day + placebo (0.9% i.v. saline). Depressive symptoms were measured using the Montgomery-Asberg Depression Rating Scale (MADRS) and the Quick Inventory of Depressive Symptomatology - Self-Report (QIDS-SR). Suicidal ideation was evaluated with the QIDS-SR item 12. Adverse psychopathological effects were measured with the Brief Psychiatric Rating Scale (BPRS)-positive symptoms, Young Mania Rating Scale (YMRS) and Clinician Administered Dissociative States Scale (CADSS). Patients were assessed at baseline, 1, 2, 4, 24 and 72 h and 7, 14, 21 and 28 days. Time to response (⩾ 50% MADRS score reduction) was the primary outcome.
By 4 weeks, more escitalopram + ketamine-treated than escitalopram + placebo-treated patients responded (92.3% v. 57.1%, p = 0.04) and remitted (76.9% v. 14.3%, p = 0.001), with significantly shorter time to response hazard ratio (HR) 0.04, 95% confidence interval (CI) 0.01-0.22, p < 0.001 and remission (HR 0.11, 95% CI 0.02-0.63, p = 0.01). Compared to escitalopram + placebo, escitalopram + ketamine was associated with significantly lower MADRS scores from 2 h to 2 weeks (peak = 3 days-2 weeks; effect size (ES) = 1.08-1.18), QIDS-SR scores from 2 h to 2 weeks (maximum ES = 1.27), and QIDS-SR suicidality from 2 to 72 h (maximum ES = 2.24). Only YMRS scores increased significantly with ketamine augmentation (1 and 2 h), without significant BPRS or CADSS elevation.
Single-dose i.v. ketamine augmentation of escitalopram was safe and effective in severe MDD, holding promise for speeding up early oral antidepressant efficacy.
Hereditary cholestasis in childhood and infancy with normal serum gamma‐glutamyltransferase (GGT) activity is linked to several genes. Many patients, however, remain genetically undiagnosed. Defects ...in myosin VB (MYO5B; encoded by MYO5B) cause microvillus inclusion disease (MVID; MIM251850) with recurrent watery diarrhea. Cholestasis, reported as an atypical presentation in MVID, has been considered a side effect of parenteral alimentation. Here, however, we report on 10 patients who experienced cholestasis associated with biallelic, or suspected biallelic, mutations in MYO5B and who had neither recurrent diarrhea nor received parenteral alimentation. Seven of them are from two study cohorts, together comprising 31 undiagnosed low‐GGT cholestasis patients; 3 are sporadic. Cholestasis in 2 patients was progressive, in 3 recurrent, in 2 transient, and in 3 uncategorized because of insufficient follow‐up. Liver biopsy specimens revealed giant‐cell change of hepatocytes and intralobular cholestasis with abnormal distribution of bile salt export pump (BSEP) at canaliculi, as well as coarse granular dislocation of MYO5B. Mass spectrometry of plasma demonstrated increased total bile acids, primary bile acids, and conjugated bile acids, with decreased free bile acids, similar to changes in BSEP‐deficient patients. Literature review revealed that patients with biallelic mutations predicted to eliminate MYO5B expression were more frequent in typical MVID than in isolated‐cholestasis patients (11 of 38 vs. 0 of 13). Conclusion: MYO5B deficiency may underlie 20% of previously undiagnosed low‐GGT cholestasis. MYO5B deficiency appears to impair targeting of BSEP to the canalicular membrane with hampered bile acid excretion, resulting in a spectrum of cholestasis without diarrhea. (Hepatology 2017;65:1655‐1669).
Brassinolides (BRs) are naturally occurring substances, which modulate plant growth and development events and have been known to improve the crop tolerance to abiotic stresses. In this study, ...possible role of exogenously applied brassinolide (BR) in alleviating the detrimental effects of drought in maize was evaluated in a rain‐protected wire‐house. Maize was subjected to drought at the start of tasseling for 6 days by withholding water application followed by foliar spray of BR (0.1 mg l−1) to assess the changes in growth, gas exchange, chlorophyll contents, protein, relative leaf water contents (RLWC), proline, malonialdehyde (MDA) and enzymatic antioxidants. Drought substantially reduced the maize growth in terms of plant height, leaf area and plant biomass. Moreover, substantial decrease in gas exchange attributes (net photosynthetic rate (A), transpiration rate (E), stomatal conductance (gs), water use efficiency (WUE), instantaneous water use efficiency (WUEi) and intercellular CO2 (Ci) was also recorded. However, exogenous application of BR remarkably improved the gas exchange attributes, plant height, leaf area, cobs per plant, seedling dry weight both under drought and well‐watered conditions. BR‐induced promotion in growth and physiological and metabolic activities were mediated through increased protein synthesis enabling maintenance of tissue water potential and activities of antioxidant enzymes lowering the lipid peroxidation under drought.
The study reported in this article attempted to obtain normative acoustic data of voice for elderly male and female speakers and to explore the educational implications of the effects of aging on ...those selected acoustic parameters. Voice samples from 21 male and 23 female elderly speakers aged 70 to 80 years were obtained on measures of 15 selected Multi-Dimensional Voice Program acoustic parameters. These data then were compared with the published norms for young and middle-aged adults. The results showed that, compared with young and middle-aged adults, elderly speakers had significantly different (usually poorer) vocal output on all of the selected acoustic parameters of voice. These findings illustrate the importance of establishing acoustic norms and thresholds for elderly men and women and stress the necessity of using discretion in making dragnostic measurements of elderly speakers' acoustic parameters of voice. This article also highlights the educational implications of such aging voice changes.
Accurate knowledge of the location and magnitude of ocean heat content (OHC) variability and change is essential for understanding the processes that govern decadal variations in surface temperature, ...quantifying changes in the planetary energy budget, and developing constraints on the transient climate response to external forcings. We present an overview of the temporal and spatial characteristics of OHC variability and change as represented by an ensemble of dynamical and statistical ocean reanalyses (ORAs). Spatial maps of the 0–300 m layer show large regions of the Pacific and Indian Oceans where the interannual variability of the ensemble mean exceeds ensemble spread, indicating that OHC variations are well-constrained by the available observations over the period 1993–2009. At deeper levels, the ORAs are less well-constrained by observations with the largest differences across the ensemble mostly associated with areas of high eddy kinetic energy, such as the Southern Ocean and boundary current regions. Spatial patterns of OHC change for the period 1997–2009 show good agreement in the upper 300 m and are characterized by a strong dipole pattern in the Pacific Ocean. There is less agreement in the patterns of change at deeper levels, potentially linked to differences in the representation of ocean dynamics, such as water mass formation processes. However, the Atlantic and Southern Oceans are regions in which many ORAs show widespread warming below 700 m over the period 1997–2009. Annual time series of global and hemispheric OHC change for 0–700 m show the largest spread for the data sparse Southern Hemisphere and a number of ORAs seem to be subject to large initialization ‘shock’ over the first few years. In agreement with previous studies, a number of ORAs exhibit enhanced ocean heat uptake below 300 and 700 m during the mid-1990s or early 2000s. The ORA ensemble mean (±1 standard deviation) of rolling 5-year trends in full-depth OHC shows a relatively steady heat uptake of approximately 0.9 ± 0.8 W m
−2
(expressed relative to Earth’s surface area) between 1995 and 2002, which reduces to about 0.2 ± 0.6 W m
−2
between 2004 and 2006, in qualitative agreement with recent analysis of Earth’s energy imbalance. There is a marked reduction in the ensemble spread of OHC trends below 300 m as the Argo profiling float observations become available in the early 2000s. In general, we suggest that ORAs should be treated with caution when employed to understand past ocean warming trends—especially when considering the deeper ocean where there is little in the way of observational constraints. The current work emphasizes the need to better observe the deep ocean, both for providing observational constraints for future ocean state estimation efforts and also to develop improved models and data assimilation methods.
Permethylation is a common derivatization method for MS-based glycomic analyses. Permethylation enhances glycan ionization efficiency in positive MS analysis and improves glycan structural stability. ...Recent biological glycomic studies have added to the growing body of knowledge and suggest the need for complete structural analysis of glycans. However, reverse phase LC analysis of permethylated glycans usually results in poor isomeric separation. To achieve isomeric separation of permethylated glycans, a porous graphitic carbon (PGC) column was used. PGC columns are well-known for their isomeric separation capability for hydrophilic analyses. In this study, we have optimized temperature conditions to overcome the issues encountered while separating permethylated glycans on a PGC column and found that the highest temperature examined, 75 °C, was optimal. Additionally, we utilized tandem MS to elucidate detailed structural information for the isomers separated. Glycan standards were also utilized to facilitate structural identifications through MS/MS spectra and retention time comparison. The result is an efficient and sensitive method capable of the isomeric separation of permethylated glycans. This method was successfully applied for the isomeric characterization of N-glycans released from the breast cancer cell lines MDA-MB-231 and MDA-MB-231BR (brain seeking). A total of 127 unique glycan structures were identified with 39 isobaric structures, represented as 106 isomers, with 21 nonisomeric glycans. Thirty seven structures exhibited significant differences in isomeric distribution (P < 0.05). Additionally, alterations in the distribution of isomeric sialylated glycans, structures known to be involved in cell attachment to the blood-brain barrier during brain metastasis, were observed.
A broiler chicken study was conducted for 42 D to evaluate their responses to 3 commercially available microbial phytases. Growth performance, nutrient digestibility, and bone mineralization at days ...21 and 42 posthatching were used as parameters of evaluation. The study was a randomized complete block design with 12 treatments, 8 replicate pens, and 25 birds per pen. Treatments included a positive control (PC), a negative control (NC) with crude protein (CP), nonphytate phosphorus (P), and calcium (Ca) reduced by 18, 1.5, and 1.8 g/kg, respectively; the NC + 4 levels of phytase A (250, 500, 750, 1,000 FTU/kg), 3 levels of phytase B (250, 500, 750 FTU/kg), and 3 levels of phytase C (500, 750, 1,000 FTU/kg). Broilers fed the NC diet had reduced (P < 0.05) performance and digestibility measures at days 21 and 42 relative to the PC. All phytase enzymes improved (P < 0.05) BW, gain, feed efficiency, and tibia ash weight and percent. Inclusion of phytase at the highest levels improved (P < 0.05) tibia ash weight by an average of 18.5 and 22% at days 21 and 42, respectively, over the NC. Phytase A linearly improved (P < 0.05) the apparent ileal digestibility (AID) of DM, Ca, P, copper, and sodium at day 21, and the AID of energy, nitrogen, and all amino acid (AA) digestibility at day 42 posthatching. Phytase B linearly (P < 0.05) improved BW gain and feed efficiency of birds at day 21 and quadratically improved (P < 0.05) the AID of nitrogen and all AA in birds at day 42. Supplementation of birds fed the NC with phytase C linearly improved (P < 0.05) the BW gain, feed intake, feed efficiency, and AID of DM, energy, nitrogen, all AA, and all minerals except manganese at day 42. In conclusion, all 3 phytase products improved the growth performance, nutrient and mineral digestibility, and bone mineralization of birds fed diets deficient in nitrogen, Ca, and P similar to or more than birds fed diet adequate in P and CP.
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