‘Sunglow’ American Witchhazel

Witchhazel (Hamamelis sp.) cultivars are now available in an array of forms and flower colors, including several native, pollinator-friendly cultivars. However, little is known about response of witchhazel cultivars to powdery mildew (Podosphaera biuncinata) or the growth and flowering characteristics of witchhazel cultivars in a nursery field production setting. To provide growth, flowering, and disease incidence data to nursery growers, a cultivar trial including 23 cultivars of witchhazel representing five species was planted Apr. 2016 in McMinnville, TN. Plant growth, flowering density, length of bloom, and foliar disease incidence were evaluated over three growing seasons between May 2016 and Oct. 2018. ‘Zuccariniana’ japanese witchhazel (H. japonica) and ‘Sunglow’ common witchhazel (H. virginiana) showed the greatest height increase during the trial, and ‘Sunglow’ also added the most width during the trial. Cultivars with negative height or width growth included Sweet Sunshine chinese witchhazel (H. mollis) and hybrid witchhazels (H. ×intermedia) Aphrodite, Twilight, and Barmstedt Gold. Ten of the 23 cultivars experienced winter injury in the form of stem necrosis. Root crown sprouts were observed for all cultivars at least once during the trial. ‘Wisely Supreme’ chinese witchhazel had the longest bloom period, followed by ‘Westerstede’ and ‘Twilight’ hybrid witchhazels, whereas ‘Quasimodo’ vernal witchhazel (H. vernalis) had the greatest density of flowers. The hybrid witchhazel cultivars Aphrodite, Nina, and Arnold Promise and the common witchhazel cultivars Green Thumb and Sunglow were resistant to powdery mildew under trial conditions in all 3 years. ‘Twilight’ and ‘Barmstedt Gold’ hybrid witchhazel, ‘Little Suzie’ common witchhazel, ‘Wisley Supreme’ chinese witchhazel, and ‘Shibamichi Red’ japanese witchhazel were moderately resistant to powdery mildew

Retinoic acid modulates chromatin to potentiate tumor necrosis factor alpha signaling on the DIF2 promoter

Transcriptional activation by nuclear hormone receptors is well characterized, but their cooperation with other signaling pathways to activate transcription remains poorly understood. Tumor necrosis factor alpha (TNFα) and all-trans retinoic acid (RA) induce monocytic differentiation of acute promyelocytic leukemia (APL) cells in a synergistic manner. We used the promoter of DIF2, a gene involved in monocytic differentiation, to model the mechanism underlying the cooperative induction of target genes by RA and TNFα. We show a functional RA response element in the DIF2 promoter, which is constitutively bound by PML/RARα in APL cells. RA stimulates release of corepressors and recruitment of chromatin modifying proteins and additional transcription factors to the promoter, but these changes cause only a modest induction of DIF2 mRNA. Co-stimulation with RA plus TNFα facilitates binding of NF-κB to the promoter, which is crucial for full induction of transcription. Furthermore, RA plus TNFα greatly enhanced the level of RNA Pol II phosphorylation on the DIF2 promoter, via synergistic recruitment of TFIIH. We propose that RA mediates remodeling of chromatin to facilitate binding of transcription factors, which cooperate to enhance Pol II phosphorylation, providing a mechanism whereby nuclear receptors interact with other signaling pathways on the level of transcription

Competition between translation initiation factor eIF5 and its mimic protein 5MP determines non-AUG initiation rate genome-wide

Citation: Tang, L., Morris, J., Wan, J., Moore, C., Fujita, Y., Gillaspie, S., … Asano, K. (2017). Competition between translation initiation factor eIF5 and its mimic protein 5MP determines non-AUG initiation rate genome-wide. Nucleic Acids Research, 45(20), 11941–11953. https://doi.org/10.1093/nar/gkx808In the human genome, translation initiation from nonAUG codons plays an important role in various gene regulation programs. However, mechanisms regulating the non-AUG initiation rate remain poorly understood. Here, we show that the non-AUG initiation rate is nearly consistent under a fixed nucleotide context in various human and insect cells. Yet, it ranges from <1% to nearly 100% compared to AUG translation, depending on surrounding sequences, including Kozak, and possibly additional nucleotide contexts. Mechanistically, this range of non-AUG initiation is controlled in part, by the eIF5-mimic protein (5MP). 5MP represses non-AUG translation by competing with eIF5 for the Met-tRNAi-binding factor eIF2. Consistently, eIF5 increases, whereas 5MP decreases translation of NAT1/EIF4G2/DAP5, whose sole start codon is GUG. By modulating eIF5 and 5MP1 expression in combination with ribosome profiling we identified a handful of previously unknown non-AUG initiation sites, some of which serve as the exclusive start codons. If the initiation rate for these codons is low, then an AUG-initiated downstream ORF prevents the generation of shorter, AUGinitiated isoforms. We propose that the homeostasis of the non-AUG translatome is maintained through balanced expression of eIF5 and 5MP

Prehospital transdermal glyceryl trinitrate in patients with ultra-acute presumed stroke (RIGHT-2): an ambulance-based, randomised, sham-controlled, blinded, phase 3 trial

Background High blood pressure is common in acute stroke and is a predictor of poor outcome; however, large trials of lowering blood pressure have given variable results, and the management of high blood pressure in ultra-acute stroke remains unclear. We investigated whether transdermal glyceryl trinitrate (GTN; also known as nitroglycerin), a nitric oxide donor, might improve outcome when administered very early after stroke onset. Methods We did a multicentre, paramedic-delivered, ambulance-based, prospective, randomised, sham-controlled, blinded-endpoint, phase 3 trial in adults with presumed stroke within 4 h of onset, face-arm-speech-time score of 2 or 3, and systolic blood pressure 120 mm Hg or higher. Participants were randomly assigned (1:1) to receive transdermal GTN (5 mg once daily for 4 days; the GTN group) or a similar sham dressing (the sham group) in UK based ambulances by paramedics, with treatment continued in hospital. Paramedics were unmasked to treatment, whereas participants were masked. The primary outcome was the 7-level modified Rankin Scale (mRS; a measure of functional outcome) at 90 days, assessed by central telephone follow-up with masking to treatment. Analysis was hierarchical, first in participants with a confirmed stroke or transient ischaemic attack (cohort 1), and then in all participants who were randomly assigned (intention to treat, cohort 2) according to the statistical analysis plan. This trial is registered with ISRCTN, number ISRCTN26986053. Findings Between Oct 22, 2015, and May 23, 2018, 516 paramedics from eight UK ambulance services recruited 1149 participants (n=568 in the GTN group, n=581 in the sham group). The median time to randomisation was 71 min (IQR 45–116). 597 (52%) patients had ischaemic stroke, 145 (13%) had intracerebral haemorrhage, 109 (9%) had transient ischaemic attack, and 297 (26%) had a non-stroke mimic at the final diagnosis of the index event. In the GTN group, participants’ systolic blood pressure was lowered by 5·8 mm Hg compared with the sham group (p<0·0001), and diastolic blood pressure was lowered by 2·6 mm Hg (p=0·0026) at hospital admission. We found no difference in mRS between the groups in participants with a final diagnosis of stroke or transient ischaemic stroke (cohort 1): 3 (IQR 2–5; n=420) in the GTN group versus 3 (2–5; n=408) in the sham group, adjusted common odds ratio for poor outcome 1·25 (95% CI 0·97–1·60; p=0·083); we also found no difference in mRS between all patients (cohort 2: 3 [2–5]; n=544, in the GTN group vs 3 [2–5]; n=558, in the sham group; 1·04 [0·84–1·29]; p=0·69). We found no difference in secondary outcomes, death (treatment-related deaths: 36 in the GTN group vs 23 in the sham group [p=0·091]), or serious adverse events (188 in the GTN group vs 170 in the sham group [p=0·16]) between treatment groups. Interpretation Prehospital treatment with transdermal GTN does not seem to improve functional outcome in patients with presumed stroke. It is feasible for UK paramedics to obtain consent and treat patients with stroke in the ultraacute prehospital setting. Funding British Heart Foundation

Interaction of the anti-apoptotic protein BAG-1 with the vitamin D receptor

BAG-1 (BCL-2 associated anti-death gene) is a multi-isoform, BCL-2 binding protein having anti-apoptotic capabilities. In addition to binding BCL-2, BAG-1 has been found to bind and regulate the function of various steroid hormone receptors. It is postulated that BAG-l's ability to inhibit apoptosis and proliferation induced by hormone receptors could lead to tumor growth if BAG-1 were to become overexpressed. In fact, BAG-1 has been found to be overexpressed in cervical and breast tumors indicating that BAG-1 may be a proto-oncogene. To further investigate BAG-1's role in steroid hormone regulation, I decided to investigate whether or not BAG-1 could bind and regulate another member of the steroid hormone receptor super family - the vitamin D 3 receptor. -- Far Western blot analysis and glutathione S-transferase -BAGp50 pull-down assays revealed that the full length 50 KDa isoform of BAG-1 could interact with the vitamin D receptor (VDR). The shorter isoforms however, could not. Gel shift assays using cell extracts from BAGp50 stably transfected U87 glioblastoma cells (U87BAG-1) showed that BAG-1 could inhibit the VDR from binding to its consenus response element, as well as a vitamin D response element (VDRE) from the P21wafl promoter. Furthermore, overexpression of BAGp50 not only resulted in an increased rate of proliferation but rendered the cells resistant to vitamin D-induced growth inhibition. A CAT construct containing a osteocalcin VDRE was employed to demonstrate that BAGp50 could also inhibit vitamin D-mediated gene expression. This was further illustrated by the fact that the presence of BAGp50 in U87 cells blocked the induction of VDR protein levels in response to vitamin D3. -- Because BAGp50 could block vitamin D3-mediated transcription of the osteocalcin VDRE it was decided to investigate whether B AGp50 could block vitamin D3-mediated upregulation of p21wafl transcription. Using p21wafl luciferase constructs, it was found that BAGp50 could inhibit vitamin D3 activation of p21wafl transcription. Interestingly, the p46 isoform of BAG-1 decreased the level of P21wafl transcription through an unknown mechanism. -- These results demonstrate for the first time that BAGp50 can bind and regulate the function of the VDR. It was also found that BAG-1 may act as a regulator of proliferation. BAGp50 and p46 could also regulate the transcription of p21wafl through two separate mechanisms, suggesting that different isoforms of BAG-1 may work together to achieve the common goal of promoting cell proliferation

A phytochemical investigation of the leaves from Erigeron philadelphicus

M.S.Leon H. Zalko

Synergy between all-trans retinoic acid and tumor necrosis factor in acute promyelocytic leukemia cell lines

Acute Promyelocytic Leukemia (APL) results from an accumulation of undifferentiated promyelocyte progenitors. Complete clinical remission of APL can be achieved through therapy with retinoic acid (RA). However, patients treated with RA alone almost invariably develop resistance to RA. We have found that tumor necrosis factor (TNF) and RA can synergize to induce differentiation of RA sensitive APL cells in a much shorter period of time than RA alone. In addition, this combination can also overcome the maturation block of RA resistant APL cells and the non-APL leukemia cell line U937 leading to differentiation. Correlating with this, we found synergistic induction of several genes at early and late time points in both the RA sensitive and resistant cell lines. Through use of neutralization antibodies, we found the protein product of one of these genes, the receptor for the macrophage colony stimulating factor, was important in mediating the differentiation effects of TNF and RA.To better understand transcriptional activation with RA and TNF we used the promoter of a gene synergistically induced by RA and TNF, Dif-2, as a model to investigate the mechanism whereby the two agents interact on the level of transcription at early time periods. We used ChIP analysis to study the accessibility of the promoter to binding by various transcription factors and the recruitment of cofactors in response to RA and TNF. We found that upon RA treatment, there was a release of corepressors and a decrease in histone methylation. This was accompanied by a subsequent increase in binding of the transcription factor PU.1, a recruitment of coactivators, as well as Snf5 (a component of the Swi/Snf complex), and an increase in histone acetylation. Interestingly, TNF could only stimulate NF-kappaB (downstream effectors of TNF) binding to the Dif-2 promoter when cells were cotreated with RA. Furthermore, TNF and RA lead to a heightened level of active, phosphorylated RNA Pol II at the Dif-2 promoter. Correlating with this was heightened recruitment of TFIIH, a known Pol II kinase. This may represent a mechanism whereby RA and TNF act in synergy to activate Pol II. These data suggest a novel mechanism for synergy between signaling pathways where RA can trigger a conformation change in a target promoter/enhancer region resulting in a more open conformation that is conducive to transcription factor binding in response to other stimuli