BA Treatment Research Articles

Phyllotaxis of vegetative shoots of Anagallis arvensis L. treated in vitro with 6-benzylaminopurine

Application of 6-benzylaminopurine (BA), a synthetic cytokinin, to the growth medium in vitro stimulates the formation of adventitious shoots on stem explants of <em>Anagallis arvensis</em>. They regenerate from dedifferentiating epidermal cells of stem internodes. The process starts from periclinal and anticlinal divisions in a few of neighbouring cells. The amount of adven-titious shoots increases with an increasing BA concentration. The BA treatment also affects phyllotaxis, but only of adventitious shoots, and not of normal lateral shoots developing in leaf axils. Leaf arrangement on adventitious shoots is always variable, even on the control medium. Generally, patterns observed in case of in vitro shoots do not differ from those reported earlier for intact <em>A. arvensis</em> plants. BA treatment also leads to the teratologic development, i.e. fasciation and the formation of abnormally shaped leaves, but again in adventitious rather than in normally developing shoots. We postulate that the variability of phyllotaxis characteristic for adventitious shoots and their responce to BA treatment, result from the fact that their apical meristems develop de novo from initially unorganised meristematic centres, whereas meristems of normally developing lateral shoots are well organised from the time of their initiation.

Hydrogen peroxide regulated photosynthesis in C4-pepc transgenic rice

In this study, we investigated the photosynthetic physiological basis in 'PC' transgenic rice (Oryza sativa L.), showing high-level expression of the gene encoding C4 phosphoenolpyruvate carboxylase (pepc), by hydrogen peroxide (H2O2). The C4-PEPC gene (pepc) from maize in the transgenic rice plants was checked by PCR. Comparison of yield components and photosynthetic indices between PC and untransformed wild-type (WT) plants indicated that increased yield in PC was associated with higher net photosynthetic rate and higher activities of phosphoenolpyruvate carboxylase (PEPC). Both PC and WT plants were treated with 1 mmol L−1 abscisic acid (ABA), 0.04% 1-butanol (BA), 2 mmol L−1 neomycin (NS), or 2 mmol L−1 diphenyleneiodonium chloride (DPI) to investigate the relationship between photosynthesis and levels of H2O2 and phosphatidic acid. In both PC and WT, ABA induced H2O2 generation and simultaneous decrease in stomatal conductance (gs). PC plants treated with BA showed decreased H2O2 content and strongly increased gs within 2 h of treatment. Similar results were observed in response to DPI treatment in PC. However, WT did not observe the decrease of H2O2 during the treatments of BA and DPI. The reduced H2O2 content in PC caused by BA treatment differed to that induced by DPI because BA did not inhibit NADPH oxidase activities. While BA induced a larger PEPC activity in PC, and higher catalase activity as well. These results indicated that the regulation of endogenous H2O2 metabolism of PC could be helpful for enhancing photosynthetic capability.

Bacillus amyloliquefaciens SC06 inhibits ETEC-induced pro-inflammatory responses by suppression of MAPK signaling pathways in IPEC-1 cells and diarrhea in weaned piglets

Enterotoxigenic Escherichia coli (ETEC) are the main pathogens which cause pro-inflammatory responses in intestinal epithelial cells and subsequent diarrhea in pigs, leading to a reduced growth rate and increased mortality. Administration of probiotics as feed additives displayed health benefits against intestinal infections. Bacillus amyloliquefaciens SC06 (Ba) is a candidate probiotic isolated from soil and kept at China Center for Type Culture Collection. In this study, we investigated the effect of Ba on the pro-inflammatory responses induced by ETEC in IPEC-1 cells and on diarrhea in the weaned piglets. The results in vitro showed that the mRNA expression of IL-6, IL-8, TNF-α, and IL-1α in IPEC-1 cells co-cultured with ETEC was significantly increased compared to the control, however, the gene expression of these genes decreased when IPEC-1 cells were incubated 24h with Ba prior to ETEC infection. Furthermore, the ETEC-mediated activation of ERK1/2, p38 and JNK in IPEC-1 cells was attenuated after Ba treatment. The results in vivo showed that the dietary supplementation with Ba increased daily body weight gain by 11.85% (P<0.05) and decreased the diarrhea incidence by 79.17% compared with the control group (P<0.01), respectively. Together, our data suggest that Ba treatment may inhibit ETEC-induced pro-inflammatory responses by suppression of MAPK signaling pathways in intestinal epithelial cells, which provides a rationale for decreased diarrhea incidence in the Ba-supplemented diet of the weaned piglets.

In vitro propagation and conservation of Diaphanoptera khorasanica (Caryophyllaceae), a threatened endemic and potential ornamental species in northeast of Iran

Tissue culture methods provide tools to supplement traditional methods for collection, propagation and preservation of endangered plant species. In this study, in vitro propagation of Diaphanoptera khorasanica Rech.f., a rare and threatened plant species with limited distribution range and population was investigated. This species has a potential as an ornamental plant. Single node explants were provided from both adult and seedling sources. Several disinfection treatments were tried to permit selection of a suitable method. Different growth regulators were used for establishment, proliferation and rooting stages. Explants showed the highest establishment percentage after 5 min treatment with 1% sodium hypochlorite (NaOCl), cultured in MS medium containing 2.2 μM 6-benzylaminopurine (BAP) and 2.4 μM indole-3-butyric acid (IBA). The highest proliferation of explants from both adult and seedling source explants was obtained from media supplemented by BA treatment in contrast to TDZ. No significant differences were found between different concentrations of BAP and TDZ. Proliferated shoots in TDZ were longer and had more internode length and less vitrification, in comparison with those in BAP. In vitro rooting of proliferated shoots just induced in liquid half-strength MS medium and rooting was not observed in solid medium. The shoots that originated from adult plants gave rise to the highest rooting rate with 4.8μM α-naphthalene acetic acid (NAA) and 2.4 μM, but NAA rooted plantlets showed higher survival percentage in acclimatization step. This study was aimed towards developing an efficient protocol for in vitro propagation of D. khorasanica and conservation of this vulnerable species.

Methyl jasmonate treatment promotes flower opening of cut Eustoma by inducing cell wall loosening proteins in petals

Establishing the technique for controlling the rate of cut flower opening is important to maintain appropriate cut flower supplies to meet consumer demand. Cut flowers of Eustoma grandiflorum (Raf.) Shinn. were held in a vase solution containing (±)-abscisic acid (ABA), 6-benzylaminopurine (BA), gibberellic acid-3 (GA), methyl jasmonate (MeJA) or 1-naphthaleneacetic acid (NAA) at 100μM. MeJA accelerated flower opening. Only the timing of flowering was earlier, and there was no change in maximum flower diameter at the fully open stage. Expansin and xyloglucan endotransglycosylase/hydrolase (XTH), regarded as cell wall loosing proteins, participate in petal growth from bud stage to the fully open stage in Eustoma. MeJA also accelerated the expression of EgEXPA2, EgEXPA3 and EgXTH1 mRNA and the accumulation of expansin and XTH protein in petals. Meanwhile, the acceleration of both flower opening and expression of these genes was not observed by ABA, BA or GA treatment. It was proposed that early flower opening by JA treatment resulted from petal cell wall loosening by accelerated expression of expansin and XTH.

Probiotic Bacillus amyloliquefaciens mediate M1 macrophage polarization in mouse bone marrow-derived macrophages

Depending on the microenvironment, macrophages can acquire distinct functional phenotypes, referred to as classically activated M1 and M2. M1 macrophages are considered potent effector cells that kill intracellular pathogens, and M2 macrophages promote the resolution of wound healing. In this study, we are interested to know whether probiotic Bacillus amyloliquefaciens (Ba) can induce macrophages polarization. Real-time fluorescence PCR analysis demonstrated that the expression of IL-1β, iNOS, TNF-α and IL-6 genes for M1 macrophages was significantly increased at 1.5 h after probiotic Ba treatment compared to the probiotic Ba-free treatment (P < 0.01), whereas the expression of M2 macrophage marker genes (Arg1, Fizz1, MR, Ym1) was decreased (P < 0.05). Furthermore, the phagocytic activity was dramatically increased in the Ba-treated BMDMs using a FITC-dextran endocytosis assay. Together, these findings indicated that probiotic Ba facilitated polarization of M1 macrophages and enhanced its phagocytic capacity. The results expanded our knowledge about probiotic function-involved macrophage polarization.

Exogenous brassinosteroids activate cytokinin signalling pathway gene expression in transgenic Arabidopsis thaliana

Transgenic Arabidopsis thaliana plants carrying the GUS reporter gene fused to the promoter of the gene of primary response to cytokinins (CKs), ARR5, were used to estimate the influence of several brassinosteroids (BRs): brassinolide (BL), epibrassinolide (EBL), homobrassinolide (HBL), and 6-o-carboxymethyloxohomocastasterone (CHC) on the expression of CK signalling genes. BRs tested differed in their ability to activate the ARR5 gene promoter in 4-day-old seedlings and 3-week-old plants. BL caused the most prominent effect, yet it was considerably less than that of 6-benzylaminopurine (BA). An increase in GUS activity was observed in both dark and light conditions; however, the rate of elevation was higher in dark conditions. The activation of the P ARR5 :GUS fusion was accompanied by a moderate induction of the P AHK :GUS constructs, in which the reporter GUS gene was fused to the promoter of one of the CK receptor histidine kinases. The effects of BL on the AHK gene promoters were organ specific and correlated with the ability of a particular AHK gene to respond to BA treatment. BL activated the AHK3 promoter in 4-day-old seedlings and in shoots and roots of 3-week-old plants without any effect in detached leaves. The AHK2 gene promoter was activated by BA and BL only in seedlings, whereas the AHK4 gene promoter was activated only in roots. BL treatment caused the coordinate elevation of the CK levels in leaves to the same degree as the activation of the P AHK :GUS construct, suggesting that the accumulation of CKs was the reason for the activation by BRs of the CK signalling genes. The data obtained provide the evidence for the involvement of BRs in the regulation of the genes of the CK signalling pathway through an increase in the CK levels. However, the exact molecular mechanisms underlying BR-induced elevation of the CK content are unclear and warrant identification in the future.

Micropropagation of self-heading Philodendron via direct shoot regeneration

The present study describes a direct shoot regeneration-based micropropagation procedure for the self-heading cultivars of Philodendron. Three types of explant (i.e. leaf lamina, petiole, stem nodal segment) were screened for their shoot induction potential following a three months treatment with 0.5mgl−1 of either 2,4-dichlorophenoxyacetic acid (2,4-D), thidiazuron (TDZ) or both. Results indicated that the leaf laminas were poor candidates for shoot induction whereas the petioles showed potential for adventitious shoot production at frequencies of 2.8–11.1% in two of the cultivars tested. Stem nodal segments were the most responsive among the three as shoots formed directly following the TDZ treatment at frequencies of 16.7–41.7% depending on the cultivar. When comparing the effectiveness of different cytokinins to induce shoot proliferation on stem nodal segments, it was found that the 0.5 and 1mgl−1 of kinetin (Kn) and 6-benzyladenine (BA) treatments resulted in higher shoot formation percentages compared to the 0.5 and 1mgl−1 of TDZ treatments in two of the three cultivars. Furthermore, more shoots were produced on BA than on Kn-supplemented media in all the three cultivars. Shoots derived from the 0.5mgl−1 of BA treatment can be induced to root following one month incubation with 0.1–1mgl−1 of indole-3-butyric-acid (IBA). The rooted shoots showed 100% survival after acclimatization in the greenhouse. The procedure reported in the present study can assist in the large-scale multiplication of elite self-heading cultivars of Philodendron in the future.

In vitro production of phenolic compounds and antioxidant activity in callus suspension cultures of Habenaria edgeworthii: A rare Himalayan medicinal orchid

An in vitro method was developed for the production of phenolic compounds from callus suspension cultures of Habenaria edgeworthii Hook. f. ex. Collett, a rare Himalayan medicinal orchid. Friable and light yellow coloured callus was induced within eight weeks when seeds were cultured on Murashige and Skoog (MS) or ½ MS medium supplemented with 1μM α-naphthaleneacetic acid (NAA). In order to increase the biomass, following another 2 weeks these calli were repeatedly subcultured (4 week interval) onto ½ MS liquid medium supplemented with 0.1μM 6-benzyladenine (BA).The effect of BA (0.0–3.0μM) and methyl jasmonate (MeJA; 10–1000μM) treatment on callus growth, total phenolic content and corresponding antioxidant activity was also determined following 4 weeks of culture on ½ MS medium (liquid). Maximum callus biomass (2.61g fresh weight) was obtained in medium containing 1.0μM BA. The total phenolic content varied significantly (p<0.05) in callus grown on BA, and ranged between 10.33 and 14.30mg gallic acid equivalent (GAE) per g dry weight (DW) (compared to control 10.17mg GAE per g DW). Antioxidant activity, measured by three in vitro assays, i.e., 2,2′- azinobis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) radical scavenging, 1,1-diphenyl-2 picrylhydrazyl (DPPH) radical scavenging and ferric reducing antioxidant power (FRAP) assays showed maximum activity in callus grown on 3.0μM BA containing medium. High performance liquid chromatography (HPLC) analysis showed the presence of high gallic acid content (143.63mg/100g DW) when compared to wild tuber (5.5mg/100g DW). The result indicate that callus grown on medium supplemented with 3.0μM BA showed high phenolic content along with increased antioxidant activity, however, callus growth was higher with 1.0μM BA. The result of this investigation has commercial applications as increased biomass production will yield active compounds, much needed by the pharmaceutical and neutraceutical industries.

6-Benzylaminopurine treatment induces increased pubescence on wheat leaves

The epidermis of wheat (Triticum aestivum L.) leaves contains trichomes that contribute to resistance to insect pests and drought tolerance. In the present study, we examined the effects of 6-benzylaminopurine (BA) and methyl jasmonate (MeJA) treatment on trichome development on the leaves of wheat cv. Norin 61 seedlings. Without phytohormone treatment, trichomes on the adaxial leaf surface were short (90 μm) and their density was low (3.6 trichomes/mm2). Both BA and MeJA treatments significantly increased the density of trichomes, and there were no significant differences between the phytohormone treatments. BA treatment increased trichome length to five times as long as that in the control, whereas MeJA treatment did not significantly affect trichome length. Since BA treatment concurrently increased the DNA content of the nuclei in trichome cells, endoreduplication of the nuclei is probably involved in trichome enlargement. These results indicate that even wheat cultivars with short trichomes retain the mechanisms for trichome enlargement and stimuli such as BA application can induce increased pubescence on wheat leaves.

Effects of Nitrogen and 6‐Benzylaminopurine on Rice Tiller Bud Growth and Changes in Endogenous Hormones and Nitrogen

ABSTRACTIn the present study, 40 mg L−1 N (NH treatment) and 20 mg L−1 6‐benzylaminopurine (6‐BA; BA treatment) were used to stimulate tiller bud growth in rice (Oryza sativa L.). The hormone changes in the tiller nodes and tiller buds and the N levels in the leaf blades, leaf sheaths, and tiller nodes were measured. The results indicate that N and 6‐BA promoted tiller bud germination and increased the indole‐3‐acetic acid (IAA) and zeatin (Z) plus zeatin riboside (ZR) levels in tiller buds and tiller nodes but decreased the abscisic acid (ABA) levels in tiller buds. Before tiller bud germination, the IAA and Z plus ZR levels increased but the ABA did not change, suggesting that IAA and Z plus ZR but not ABA may be the key regulators of tiller bud growth. From 4 d posttreatment, the tiller bud growth of the BA‐treated plants markedly slowed but the buds of the NH‐treated plants continued to grow quickly. We also found that the N levels in the leaf blades, leaf sheaths, and tiller nodes of the NH‐treated plants increased significantly as compared to those of the control plants (the plants treated with 10 mg L−1 N). However, no significant differences in N levels were observed between the plants of BA and control treatments. This study suggests that N may regulate tiller bud growth in two ways: by regulating N metabolism and regulating endogenous hormones. External 6‐BA promoted tiller bud germination by regulating endogenous hormones, but tiller bud growth was not maintained because it could not increase the N levels in the plants.

Sodium butyrate induces the production of cyclooxygenases and prostaglandin E2 in ROS 17/2.8 osteoblastic cells

ObjectiveSodium butyrate (butyric acid; BA) is a major metabolic by-product of the anaerobic periodontopathic bacteria present in subgingival plaque. We examined the effects of BA and/or indomethacin on cell proliferation, the expression of cyclooxygenases (COXs), prostaglandin (PG) receptors (EP1-4), extracellular matrix proteins, such as type I collagen and osteopontin, and PGE2 production, using ROS17/2.8 cells as osteoblasts. MethodsThe rat clonal cell line ROS 17/2.8 was cultured with 0, 10−5, 10−4, and 10−3M BA in the presence or absence of 0.5μM indomethacin, for up to 7 days. The expression of COX-1, COX-2, EP1, EP2, EP3, EP4, type I collagen, and osteopontin was examined at the mRNA and protein levels using real-time PCR and Western blotting, respectively. The amount of PGE2 in the culture medium was measured by ELISA. ResultsProliferation of ROS 17/2.8 cells was not affected by the addition of BA. However, PGE2 production and the expression of COX-1 and COX-2 increased with the addition of BA. In contrast, indomethacin, an inhibitor of COX, blocked the stimulatory effect of BA. Furthermore, EP2 expression increased with BA treatment, whereas EP1 expression was not affected and the expression of EP3 and EP4 was not detected. The addition of BA also increased the expression of type I collagen and osteopontin. Indomethacin blocked about 50% of the stimulatory effect of BA on type I collagen, whereas it did not block the effect on osteopontin. ConclusionsThese results suggest that BA induces PGE2 production by increasing the expression of COX-1 and COX-2 in osteoblasts, and that an autocrine action of the produced PGE2, via EP1 or BA-induced EP2, is related to an increase in type I collagen expression by BA.

Sumoylation of Sp1 is increased by betulinic acid and led to RNF‐mediated degradation

Our recent findings show that Sp1 can accumulate during tumorigenesis after it undergoes posttranslational modification, namely, sumoylation, and phosphorylation. We found that Sp1 was the substrate of the ubiquitin E3 ligase, RNF4. Mapping of the domain involved in the interaction between Sp1 and RNF4 revealed 2 regions that interacted with each other. One was a SUMO‐modifier in the N‐terminus of Sp1(Lys16) that interacted with the SUMO‐interacting motif (SIM) of RNF4, and the other was the C‐terminus of Sp1 that interacted with RNF4 in a SIM independent manner. We also found that a previous reported anticancer drug‐betulinic acid (BA) increases Sp1 sumoylation, facilitating interaction between Sp1 and RNF4, which leads to Sp1 degradation through the ubiquitin‐dependent proteasome pathway. Here, we elucidate that Sp1 degradation through BA treatment facilitate sumoylation of Sp1 during cancer therapies.

Fully Human Anti-Hepcidin Antibodies Mobilize Iron and Treat Inflammatory Anemia in a Mouse Model.

Abstract 626Iron maldistribution has been associated with multiple diseases including atherosclerosis, diabetes, neurodegenerative disorders and the anemia of inflammation (AI). Hepcidin is a 25aa peptide that is the central mediator of iron homeostasis. It is produced in response to inflammation and is implicated in limiting the response to erythropoietin (EPO) even in the presence of adequate body iron stores. To further explore the role of hepcidin in inflammatory anemia, a novel EPO-resistant model of AI was developed in mice and the effects of a high affinity fully human anti-hepcidin antibody (12B9m; 2pM affinity) were examined.Since antibody 12B9m had low affinity for mouse hepcidin (10-20nM), human hepcidin (hHepc) knock-in mice were created. Strains were generated with either the mouse hepc1 gene or both hepc1 and 2 genes replaced with the single hHepc gene (HAMP). To induce inflammation and establish AI, mice were injected intraperitoneally with heat-killed Brucella abortus (BA: 3×108 particles/mouse): a procedure adapted from an inflammatory fatigue model. BA treatment led to increased serum levels of IL-6 (3818 ± 548 pg/ml versus <10 pg/ml in controls, p<0.0001) and hepcidin (216 ± 9 ng/ml versus 99 ± 6 ng/ml, p<0.0001) by 6 hours. Hypoferremia reached a nadir 3 days after BA treatment (total serum iron was 201 ±10 mg/dL versus 370 ± 19 mg/dL in controls, p<0.0001). Mice developed anemia, with hemoglobin (Hb) reaching a nadir 10-14 days after BA and returning to normal by day 28. Treatment with EPO (300 mg/kg 8 days after BA) had no effect on hemoglobin by day 14 (7.6 ± 0.4 g/dL in controls and 7.6 ± 0.5 g/dL with EPO; NS).Treatment with antibody 12B9m (5mg/mouse; n=6) increased serum iron at day 3 (907 ± 47 mg/dL versus 201 ± 10 mg/dL, p<0.0001) in AI mice, and restored response to EPO (day 14 Hb of 10.0 ± 0.4 g/dL versus 7.6 ± 0.5 g/dL for control antibody treatment; p<0.001, n=5). Early iron-dependent red cell parameters such as reticulocyte MCV were also increased (71.5 ± 5.2 fL versus 56.7 ±1.9 fL; p<0.05, n=5/group), presumably due to the increased iron availability. In a larger study, 12B9m treatment alone (without EPO co-administration) led to a significant increase in Hb (10.0 ± 0.5 g/dL versus 8.0 ± 0.3 g/dL for control antibody treatment; p<0.001, n=14/group).Treatment with 12B9m did not affect inflammatory cytokine induction (e.g., IL-6 levels at 6 hours post-BA of 4996 ± 487 pg/ml versus 3818 ± 548 pg/ml in the control antibody group: NS), indicating that neutralization of hepcidin alone was sufficient to allow an effective response to EPO.These data indicated that antibodies which neutralize hepcidin may treat AI by increasing serum iron availability for erythropoiesis. The ability to manipulate iron uptake and redistribution in vivo may also offer promise in other diseases where iron maldistribution is involved. Disclosures:Cooke:Amgen: Employment. Augustic:Amgen: Employment. Sloan:Amgen: Employment. Tran:Amgen: Employment. Arvedson:Amgen: Employment. Juan:Amgen: Employment. Sasu:Amgen: Employment.

Barium Toxicity Effects in Soybean Plants

Barium (Ba)-induced phytotoxicity at 100, 1000, or 5000 microM Ba in soybean plants (Glycine max) was investigated under hydroponic culture conditions. Soybean growth and leaf photosynthetic activity were significantly inhibited by all three levels of Ba treatments. In the case of photosynthetic activity, 5000 microM Ba treatment shutdown stomatal opening and perturbed carbon fixation metabolism and translocation. However, 100 and 1000 microM Ba treatments shut down stomatal opening and inhibited carbon fixation, but without perturbation of leaf carbon fixation-related metabolism. Potassium (K) absorption by soybean roots was also reduced in all three Ba treatments. This decreased K absorption reduced K localization at guard cells. Barium accumulation in guard cells also inhibited K transport from epidermal cells to guard cells. This lack of K in guard cells resulted in stomatal closure. As a result of inhibition of K transport into guard cells and stomatal shutdown, photosynthetic activity and plant productivity were inhibited. Our experiment indicates that Ba has phytotoxic effects on soybean plants by inhibiting photosynthesis.

The effect of beta-alanine supplementation on neuromuscular fatigue in elderly (55–92 Years): a double-blind randomized study

BackgroundAgeing is associated with a significant reduction in skeletal muscle carnosine which has been linked with a reduction in the buffering capacity of muscle and in theory, may increase the rate of fatigue during exercise. Supplementing beta-alanine has been shown to significantly increase skeletal muscle carnosine. The purpose of this study, therefore, was to examine the effects of ninety days of beta-alanine supplementation on the physical working capacity at the fatigue threshold (PWCFT) in elderly men and women.MethodsUsing a double-blind placebo controlled design, twenty-six men (n = 9) and women (n = 17) (age ± SD = 72.8 ± 11.1 yrs) were randomly assigned to either beta-alanine (BA: 800 mg × 3 per day; n = 12; CarnoSyn™) or Placebo (PL; n = 14) group. Before (pre) and after (post) the supplementation period, participants performed a discontinuous cycle ergometry test to determine the PWCFT.ResultsSignificant increases in PWCFT (28.6%) from pre- to post-supplementation were found for the BA treatment group (p < 0.05), but no change was observed with PL treatment. These findings suggest that ninety days of BA supplementation may increase physical working capacity by delaying the onset of neuromuscular fatigue in elderly men and women.ConclusionWe suggest that BA supplementation, by improving intracellular pH control, improves muscle endurance in the elderly. This, we believe, could have importance in the prevention of falls, and the maintenance of health and independent living in elderly men and women.

Abstract 2163: Long-term Follow-up (&gt;3 Years) of Patients Treated With Sirolimus-Eluting Stents for In-stent Restenosis. Results of a Randomized Study.

Background: The value of drug-eluting stents in patients (P) with in-stent restenosis (ISR) has been established. However, the long-term results of this strategy in P with ISR remains unknown. Objective: We sought to determine the long-term clinical outcome of P treated with sirolimus-eluting stents (SES) for ISR. Methods: A systematic, pre-specified, long-term clinical follow-up (FU) was performed in all P included in the RIBS II (Restenosis Intra-stent: Balloon angioplasty [BA] vs elective SES implantation) randomized trial. In RIBS II 150 P with ISR after bare-metal stent implantation were included: 74 allocated to BA and 76 to SES. Late angiography was obtained in 96% of eligible P. A structured clinical questionnaire (cardiac/non cardiac death, myocardial infarction [MI], target vessel revascularization [TVR], thrombosis [TH], and medical therapy) was used during FU. Results: Angiographic restenosis (primary end-point) was more frequently found in the BA arm (39% vs 11%, p&lt;0.001). Clinical FU at 1-year was obtained in 150 P (100%). During this time period 6 P died (3 SES, 3 BA), 4 P suffered a MI (2 SES, 2 BA), 2 P experienced TH (1 P in each arm) and 30 required TVR (8 SES, 22 BA, p&lt;0.01). A complete clinical FU &gt;3 years was obtained in 145 P (97%) (mean 38±9 months, median 40 months [IQR 37–42]). Late events (after 1 year, non-exclusive) included: 3 deaths (1 SES, 2 BA), 3 MI (3 SES, 2 due to late TH) and 7 late TVR (5 SES, 2 BA). At 4 years, event-free survival was 76% in the SES arm and 65% in the BA arm (p=0.03). Survival free from TVR at 4 years was 80% in the SES arm and 67% in the BA arm (p=0.02). Conclusion: In P with ISR SES implantation improve the long-term clinical outcome as compared with BA treatment.

The Effect of Indole-3-Acetic Acid and Benzyladenine on Sequential Leaf Senescence on Helianthus Annuus L. Seedlings

ABSTRACTThe effect of IAA and BA on the mechanism of senescence which occurs in the cotyledons of Helianthus annuus L. seedlings showing epigenic germination has been investigated. In this study 0,01μM and 10μM IAA solutions were sprayed to the leaves and BA solutions with the same concentrations—to the cotyledons of 16 days-old seedlings. We have determined chlorophyll content, protein amount, peroxidase activity and the changes in the membrane permeability in the cotyledons of the treated and control seedlings. Compared to the control cotyledons IAA in the concentrations of 10μM and 0,01μM caused a breakdown of chlorophyll at the ratio of 64% and 54%, respectively. Soluble protein levels in the IAA at 10μM treatment decreased in the ratio of 12%. IAA treatments with 10μM and 0,01μM stimulated POD activity with about 11 and 8 times, respectively. On the other hand, it has been observated that 10μM and 0,01μM BA treatment retarted senescence, whereas 0,01μM BA and 0,01 μM IAA when were applied together enhanced senescence. It has been discussed that IAA which is known as an allosteric activators of cytokinin oxidase stimulates the cytokinin oxidations so it increases senescence that occurs in the cotyledons. In this research, according to the results it has been thought that the senescence signal may be a substance in the nature of IAA.

A Randomized Comparison of Sirolimus-Eluting Stent With Balloon Angioplasty in Patients With In-Stent Restenosis: Results of the Restenosis Intrastent: Balloon Angioplasty Versus Elective Sirolimus-Eluting Stenting (RIBS-II) Trial

We sought to assess the effectiveness of sirolimus-eluting stents (SES) in patients with in-stent restenosis (ISR). Treatment of patients with ISR remains a challenge. The Restenosis Intrastent: Balloon Angioplasty Versus Elective Sirolimus-Eluting Stenting (RIBS-II) study is a multicenter randomized trial conducted in 150 patients with ISR (76 allocated to SES and 74 to balloon angioplasty [BA]). The primary end point was recurrent restenosis rate at nine months. Secondary end points included prespecified subgroup analysis, lumen volume on intravascular ultrasound (IVUS), and a composite of major clinical events at one year. Angiographic success was obtained in all patients. At 9-month angiographic follow-up (96% of eligible patients) minimal lumen diameter was larger (2.52 mm [interquartile range (IQR) 2.09 to 2.81] vs. 1.54 mm [IQR 0.91 to 2.05]; p < 0.001) and recurrent restenosis rate was lower (11% vs. 39%; p < 0.001) in the SES group. Prespecified subgroup analyses were consistent with the main outcome measure. Lumen volume on IVUS at 9 months was also larger (279 mm3 [IQR 227 to 300] vs. 197 mm3 [IQR 177 to 230]; p < 0.001) in the SES group. At one-year clinical follow-up (100% of patients), the event-free survival (freedom from death, myocardial infarction, and target vessel revascularization) was significantly improved in the SES group (88% vs. 69%; p < 0.004) as the result of a lower requirement for target vessel revascularization (11% vs. 30%; p < 0.003). In patients with ISR, the use of SES provides superior long-term clinical, angiographic, and IVUS outcome than BA treatment.

Fire disturbance disrupts co‐occurrence patterns of terrestrial vertebrates in Mediterranean woodlands

AbstractAim This paper uses null model analysis to explore the pattern of species co‐occurrence of terrestrial vertebrate fauna in fire‐prone, mixed evergreen oak woodlands.Location TheErico–Quercion ilicisof the Mediterranean belt (50–800 m a.s.l.) in the Madonie mountain range, a regional park in northern Sicily (37°50′ N, 14°05′ E), Italy.Methods The stratified sampling of vertebrates in a secondary succession of recent burned areas (BA, 1–2 years old), intermediate burned areas (INT, 4–10 years old) and ancient burned areas (CNB, &gt; 50 years old), plus forest fragments left within burned areas (FF, 1–2 years old) permitted the comparison of patterns of species co‐occurrence using a set of separate presence/absence matrices. First, the breeding avifauna derived from standardized point counts was analysed using Stone &amp; Roberts’C‐score, and by a null model algorithm (fixed/equiprobable). Secondly, the analysis was repeated using all vertebrate species recorded in the succession.Results Sixty‐five species were recorded in the 2‐year study period in the four sample treatments. Birds were found to make up the largest component (63%) of the recorded assemblage. The BA treatment had the lowest species richness, followed in order by the small, medium and large FFs, and then by the CNBs. For both analyses (birds and total vertebrates), theC‐scores were quite small and not significantly different from those that could be expected by chance in the BA and INT burned areas; this indicates a random co‐occurrence among vertebrates of those assemblages. Contrariwise, for both analyses in the CNBs, theC‐scores were large and significantly different from the simulated indices, thereby indicating a non‐random co‐occurrence pattern (segregation) of vertebrates in the undisturbed woodlands. In addition,C‐score values for the surviving FFs show a significant aggregation of species.Main conclusions The null model analyses highlighted a new aspect of fire disturbance in Mediterranean woodland ecosystems: the disruption in patterns of co‐occurrence in the terrestrial vertebrate community. Wildfire alters community organization, inducing, for at least 10 years, a random aggregate of species. Communities re‐assemble themselves, showing the occurrence of species segregation at least 50 years after fire.