Sesquiterpene hydrocarbons (C15) not only share the structural properties thought to lend protective qualities to isoprene and monoterpene hydrocarbons, but also react rapidly with ozone, suggesting that sesquiterpenes may similarly enhance tolerance of abiotic stresses. To test whether sesquiterpenes protect plants against ozone, UVB light, or drought, we used transgenic lines of the wild tobacco Nicotiana attenuata. The transgenic plants expressed a maize terpene synthase gene (ZmTPS10) which produced a blend of (E)-ß-farnesene and (E)-α-bergamotene, or a point mutant of the same gene (ZmTPS10M) which produced (E)-ß-farnesene alone,. (E)-ß-farnesene exerted a local, external, and transient ozone-quenching effect in ozone-fumigated chambers, but we found no evidence that enhanced sesquiterpene production by the plant inhibited oxidative damage, or maintained photosynthetic function or plant fitness under acute or chronic stress. Although the sesquiterpenes (E)-ß-farnesene and (E)-α-bergamotene might confer benefits under intermittent heat stress, which was not tested, any roles in relieving abiotic stress may be secondary to their previously demonstrated functions in biotic interactions.Among the terpenes, isoprene (C Supernatants were evaporated to 250 μL in a rotary evaporator and analyzed using an Agilent 1200 HPLC/MS/MS. Sample (5 μL) was injected and eluted with a flow rate of 800 μL/min through an Agilent Zorbax Eclipse DBC18 1.8 μm column. A gradient of 0.05% formic acid and acetonitrile (ACN) began with 5% ACN from 0–0.5 min and progressed linearly to 100% ACN at 4.5 min followed by washing with 100% ACN from 5–5.5 min and 3 min re-initialization at 5% ACN. SA and ABA were quantified relative to the labeled internal standards using MS/MS monitoring for the ion pairs 140.9/97.0 ([2H4] SA), 136.8/93.1 (SA), 269/159.2 ([2H6]ABA), and 263/153.2 (ABA) as in Wang et al. [Frozen leaf tissue (100 mg) was extracted overnight at -20°C with 1.5 mL methanol containing the labeled internal standards [g et al. .For each experiment , treatment effects and genotype-treatment interactions were tested by two-way ANOVA using SPSS . Phytohormone data were log-transformed and moisture data arcsine-transformed for homoscedasticity. Complete statistical results are documented in E)-β-farnesene with ozone, which has been observed in smog chambers [E)-β-farnesene into empty chambers containing ca. 500 ppb ozone lowered within-chamber ozone concentrations by >100 ppb within a few minutes (E)-β-farnesene from chambers containing 350 ppb ozone (E)-β-farnesene.We tested whether the reaction of -β-farnesene-mediated ozone destruction in the leaf headspace would protect plants, we supplemented the ozone-susceptible N. tabacum cv. BelW3 with external (E)-β-farnesene diluted in acetonitrile and impregnated into cotton tampons. A dose of 2000 μg plant-1 h-1 visibly reduced ozone-induced foliar necrosis (-1 h-1 was not protective (E)-β-farnesene from the 2000 μg plant-1 h-1 treatment reached and was retained in the leaf, although the 10-fold greater leaf retention in the control fumigation suggested that most (E)-β-farnesene was destroyed in the headspace -β-farnesene with ozone could protect plants by destroying ozone in the headspace around leaves. Yet whether endogenous (E)-β-farnesene production would be sufficient to achieve leaf protection, either by acting in the headspace or within the leaf itself, remained unclear. To test for benefits of endogenous production, we transformed N. attenuata with two versions of a maize sesquiterpene synthase expressed behind a constitutive 35S promoter. The enzyme ZmTPS10 (TPS10 line) yielded nearly equal amounts of (E)-β-farnesene and (E)-α-bergamotene, whereas the point mutant Zmtps10M produced primarily (E)-β-farnesene (E)-β-farnesene: t(2.29) = 1.11, p = 0.36; (E)-α-bergamotene: t(2.35) = 1.21, p = 0.33). Emission of the TPS10- and TPS10M-catalyzed sesquiterpenes from the transformed N. attenuata lines was approximately twice as high as the sesquiterpene emission rate measured from herbivore-damaged maize plants, and exceeded undamaged plant emission by an order of magnitude [External supplementation indicated that the reaction of  = 734.3, p<0.001) and significant decreases in moisture content  = 25.95, p<0.001)) and photosynthetic rate  = 7.81, p<0.016), all common effects of ozone exposure [F = 0.88, p = 0.37; moisture: F = 0.07, p = 0.80; photosynthetic rate: F = 0.04, p = 0.85). Surprisingly, ozone treatment had no significant effect on three markers of oxidative stress in leaves: malondialdehyde (MDA), a product of lipid peroxidation  = 0.55, p = 0.47); oxidative radical absorbance capacity (ORAC)  = 0.49, p = 0.50); and rutin  = 0.16, p = 0.69), a flavonoid glycoside that dominates the leaf phenolic profile of N. attenuata and whose induction plays a role in UVB tolerance [Sesquiterpene synthase-transformed and WT reatment . Ozone-fexposure . A two-wolerance . A TPS10E)-β-farnesene’s reactivity and protective potential from the ozone experiments with BelW3, and we hypothesized that protection against UVB exposure would accrue from within-leaf terpenes, we used the TPS10M line with the highest leaf (E)-β-farnesene content . To control for effects of transformation unrelated to sesquiterpenes, we included a second, independently transformed TPS10M line .To directly test for within-leaf antioxidant effects of sesquiterpenes, we exposed sesquiterpene synthase-transformed and WT plants to UVB irradiation. Since we had evidence of  = 12.73, p = 0.001), while chlorophyll fluorescence  = 55.20, p<0.001) and photosynthetic rate  = 24.88, p<0.001) were significantly decreased, but no significant differences emerged between responses of TPS10M and WT plants  = 0.62, p = 0.55; chlorophyll fluorescence: F = 0.79, p = 0.47; photosynthetic rate: F = 0.42, p = 0.66). MDA measurements did not show a significant treatment effect  = 2.45, p = 0.14). The daily UVB exposure was continued for a total of 8 weeks, leading to significant decreases in height  = 157.00, p<0.001) and seed capsule number  = 38.15, p<0.001), but again, no differences in responses to treatment appeared between genotypes  = 0.50, p = 0.61; seed capsule number: F = 0.16, p = 0.86).The morphological responses of both TPS10M and WT leaves, which assumed a curled appearance and crispy texture, testified to the severity of the UVB exposure . After 8E)-β-farnesene-mediated resistance to drought stress using the same two TPS10M lines used in the UV experiment. Within 3 d of treatment onset, all drought-treatment plants raised their leaves, a sign of water stress in N. attenuata. At the time of tissue harvest, 6 days into the treatment, abscisic acid (ABA) levels were double those in controls  = 13.39, p = 0.001) and photosynthesis was decreased by 80%  = 562.33, p<0.001), but responses did not differ between TPS10M and WT  = 0.60, p = 0.56; photosynthesis: F = 0.12, p = 0.89). Although shoot MDA did not show a treatment effect  = 1.07, p = 0.31), root MDA increased  = 12.57, p = 0.001), but to a similar extent in all lines  = 0.71, p = 0.50). Ion leakage also increased  = 8.26, p = 0.007). Increases tended to be less dramatic in the (E)-β-farnesene emitter TPS10M1 than in WT, but TPS10M2 was no different from WT in its response, nor was the genotype x treatment term significant in an ANOVA  = 1.53, p = 0.23). Total fresh mass after 6 d of treatment  = 50.86, p<0.001), plant height after 2 weeks of treatment  = 152.19, p<0.001), and seed capsule count after an 11 d drying period in both treatment groups  = 16.94, p<0.001) all showed significant declines with drought, but no differences between the responses of TPS10M and WT  = 1.72, p = 0.20; height: F = 0.53, p = 0.59; seed capsule number: F = 1.06, p = 0.35).We tested for (5) and monoterpene (C10) hydrocarbons have been demonstrated to protect plants from oxidative stress. We tested whether sesquiterpene (C15) hydrocarbons, many of which react rapidly with ozone, also confer such protection when externally supplemented or internally generated. After demonstrating the ozone-quenching properties of the sesquiterpene (E)-ß-farnesene in empty fumigation chambers ((E)-ß-farnesene protected plants of an ozone-sensitive N. tabacum cultivar, BelW3, from visible cell death symptoms of ozone exposure ((E)-ß-farnesene was not protective. To test for benefits of endogenous sesquiterpene production, we used N. attenuata transformed with a Z. mays sesquiterpene synthase gene (35S::TPS10) that produced an (E)-ß-farnesene and (E)-α-bergamotene blend, or a point mutant of the same gene (35S::TPS10M) that produced primarily (E)-ß-farnesene. Augmenting endogenous sesquiterpene production by engineering these genes into N. attenuata did not enhance tolerance to oxidative stress under conditions of ozone, UVB, or drought.Plant-produced isoprene -ß-farnesene to quench headspace ozone (-1 h-1 conferred visible benefits (Z. mays [Solanum tuberosum [(E)-ß-farnesene (20 μg plant-1 h-1) was not protective ((E)-ß-farnesene recovered from supplemented, ozone-fumigated leaves -ß-farnesene and (E)-α-bergamotene, in the ecological model plant Nicotiana attenuata. The transformants grew and developed normally and had no significant physical or chemical differences from WT besides emission of the target volatiles [(E)-ß-farnesene recovered from leaves of BelW3plants externally supplemented with 2000 μg/plant ((E)-ß-farnesene were mediated by within-leaf activity.External supplementation experiments do not mimic the within-leaf sesquiterpene distribution resulting from olatiles . Sesquitolatiles . Becauseμg/plant , our train planta benefit. Based on the mechanisms proposed for sesquiterpene function, sesquiterpenes can be hypothesized to reduce levels of reactive oxygen species (ROS), especially in lipophilic cellular compartments, and thereby reduce oxidative damage and alter ROS-mediated signaling cascades [N. attenuata [Given the demonstrated reactivity of sesquiterpenes with ozone, we expected that sesquiterpene production might reduce ozone concentrations in the leaf boundary layer, as suggested for isoprene , or provcascades . Visiblettenuata ; shorterE)-β-farnesene emitted at 20 μg plant-1 h-1 had a short lifetime in the chamber headspace and did not confer benefits to neighboring plants. Diffusion effects [It is possible that the proximity of WT and transgenic plants in the fumigation chambers could have enabled WT to benefit from transgenic neighbors’ emissions. However, our experiments with BelW3 showed that -ß-farnesene emission increases in a number of Pinus species in response to high light levels [Z. mays grown under high light [(E)-ß-farnesene was protective against the effects of UVB. UVB treatment strongly affected leaf morphology -ß-farnesene is stimulated by herbivory and leaf excision [E)-β-farnesene conferred no protection against this stress at either the chemical or whole-plant level: the effects of drought on ABA accumulation, electrolyte leakage, photosynthetic rate, and reproductive fitness correlates were similar in N. attenuata TPS10M and WT plants.If sesquiterpenes can diffuse across membranes and quench reactive oxygen species artments . Correlaartments . Althougartments ,58, in w winters . In Citrt stress . In Z. mexcision , which iexcision . In our E)-β-farnesene.Although our results did not demonstrate that sesquiterpenes ameliorate oxidative stress, they also do not directly refute the antioxidant hypothesis for terpene-mediated abiotic stress tolerance , since tOur results also do not directly challenge the membrane stabilization-mediated thermotolerance hypothesis . SharkeyA. thaliana, but this conclusion has been questioned [Z. mays [Spinacea olearacea thylakoids [Ours is not the first study to report that terpene supplementation does not increase tolerance to abiotic stress, but our study withstands the criticisms directed at earlier studies that showed a lack of benefit from added isoprene. Loivamäki and colleagues  found noestioned  because [Z. mays . Isoprenylakoids  or increylakoids , but theN. attenuata. This wild tobacco has weathered extremes of temperature, drought, and light stress over its evolutionary history in the deserts of the American southwest [N. tabacum, might facilitate detection of terpene function in abiotic stress resistance. Genetic transformation of N. tabacum has previously revealed a small but significant role of isoprene production [A subtle protective influence of sesquiterpenes might be undetectable against the stress-adapted background of outhwest  and has outhwest . Using moduction  in toleroduction , and mig(E)-ß-farnesene and (E)-α-bergamotene in resistance to abiotic stress. Previous work suggests that these two sesquiterpenes may instead function against biotic stresses. ZmTPS10 transformation of A. thaliana attracted the parasitic wasp, Cotesia marginiventris, to its Lepidopteran host [(E)-α-bergamotene was emitted from N. attenuata in response to herbivore-mediated jasmonate signaling [N. attenuata’s herbivores [ZmTPS10-produced sesquiterpenes may also primarily mediate biotic interactions in their native maize, where emission is more responsive to herbivory than to light, temperature, and humidity [(E)-ß-farnesene and (E)-α-bergamotene in biotic interactions [Our results argue against utility of ran host . Similarignaling  and attrrbivores . ZmTPS10humidity . Bioassaractions .A. thaliana plants under heat stress [N. attenuata, such signaling seems unlikely, since TPS10 and TPS10M transformants were chemically indistinguishable from WT and did not influence the defense physiology of WT neighbors when the two genotypes were grown together in the same pot [Another role for sesquiterpenes might be as within- or between-plant signals. The enhanced growth of isoprene-emitting t stress , and thet stress  or leaf t stress , would bt stress  or primet stress . In N. asame pot . Effects(E)-ß-farnesene and (E)-α-bergamotene, including the highly ozone-reactive α-humulene or (E)-β-caryophyllene [Sesquiterpenes other than phyllene , might bphyllene , a diverS1 Fig(DOCX)Click here for additional data file.S2 Fig(DOCX)Click here for additional data file.S3 Fig(DOCX)Click here for additional data file.S1 Table(XLSX)Click here for additional data file.


In a subgroup, advanced HPs  of a medium-sized teaching hospital. Adult patients with severe sepsis were included and received standard goal-directed therapy (Surviving Sepsis Guidelines). Every patient received an arterial line and a central venous line in the upper diaphragm position. A subgroup received pulse contour cardiac output (PiCCO)-guided resuscitation and PT measurements. Pearson correlation coefficients (PCCs) were calculated between HPs and SL, which were measured every 4 hours for the first 48 hours after inclusion. n = 11) and pneumosepsis (n = 7). Mean HPs (with SD and range) were respectively: MAP 73 mmHg , HR 101 beats/minute , CVP 12 mmHg , UP 55 ml/hour , SL 3.2 mmol/l , CI 4.1 ml/kg/minute , GEDI 871 ml/m2 , ELWI 11 ml/kg , PT 32.1 C , and SvO2 75% . Relevant and significant (P < 0.005) PCCs between HPs and SL were respectively: MAP -0.417, HR 0.195, UP −0.237, SvO2 −0.204 and PT −0.569. Figure Twenty-five patients 12 men) were included. Mean age was 68 years (30 to 93), mean APACHE II score 31 (20 to 42). The most frequent reasons for IC admission were abdominal sepsis (2 men wer2 are significantly correlated to levels of SL, but clinical value might be limited due to the relatively low correlation coefficients. In a small subgroup, PT is better correlated to the level of SL.The conventional HPs MAP, HR, UP and SvO


Association of two HLA class I variants with HIV-1 pretreatment viremia, CD4+ T cell count at the care-entry and CD4+ T cell nadir.414 HIV-positive Caucasians (30% women) aged 19-73 years were genotyped for HLA-C -35 (rs9264942) and HLA-B*5701 variants. HIV-1 viral load, as well as CD4+ T cell count at care-entry and nadir, were compared across alleles, genotypes and haplotypes.HLA-C -35 C/C genotype was found in 17.6% patients, C/T genotype in 48.1%, and T/T genotype in 34.3% patients. HLA-B*5701 variant was present in 5.8% of studied population. HIV plasma viremia in the group with C allele was significantly lower (p=0.0002) compared to T/T group , while CD4+ T cell count at baseline was notably higher among C allele carriers compared to T/T homozygotes  (p=0.0007). Moreover, CD4+ T cell nadir among patients with C allele [median: 205 (IQR:83.5-390) cells/μl] was significantly higher compared to T/T group [median: 133 (IQR:46-328) cells/μl] (p=0.006). Among cases with HLA-B*5701 allele, significantly lower pretreatment viremia and higher baseline CD4+ T cell count were found  compared to HLA-B*5701 negative individuals. The lowest viremia (mean: 3.85 log [SD:1.3]) HIV-RNA copies/ml and the highest baseline and nadir CD4+ T cell [median: 476 (IQR:304-682) vs. median: 361 (IQR: 205-574) cells/μl, respectively) were found in individuals with HLA-B*5701(+)/HLA-C –35 C/C haplotype.HLA-C -35 C and HLA-B*5701 allele exert a favorable effect on the immunological (higher baseline and nadir CD4+ T cell count) and virologic  variables. This protective effect is additive for the compound HLA-B*5701(+)/HLA-C -35 C/C haplotype. Human leukocyte antigens C (HLA-C) strongly influence immunological activity in chronic viral infections  as well as in autoimmune diseases e.g. Crohn’s disease, autoimmune liver diseases, Graves’ disease and psoriasis vulgaris ,2,3,4,5.HLA-C antigens play an important role in HIV control through two mechanisms: acting as ligands for killer immunoglobulin-like receptors (KIRs) presented on natural killer (NK) cells and directly by antigen  presentation to cytotoxic T cells ,6,7.inter alia, on the level of HLA-C expression  (p = 0.0007) .Similarly, the CD4+ T cell nadir among patients with at least one HLA-C -35 C allele was significantly higher compared to T/T homozygotes [median: 205 (IQR: 84–390) cells/μl vs. median: 133 (IQR: 46–328) cells/μl] (p = 0.006) .In patients with HLA-B*5701variant, significantly lower pretreatment viral load and higher CD4+ T cell count at baseline were observed in comparison with HLA B*5701 negative group  were found in the group with the compound HLA-B*5701(+)/HLA-C locus -35 C/C haplotype Figs  and 2f.Associations between HLA-B*5701 and HLA-C -35 rs9264942 variants, HIV-1 viral load, baseline CD4+ T cell count and nadir were also analyzed separately for the HIV/HCV coinfected and HIV monoinfected groups. Results were in line with the findings presented for the entire group. In the HIV/HCV coinfected patients, lower pretreatment viral loads were observed for the HLA-C -35 C allele compared to T/T homozygotes [mean: 4.4 (SD: 1.0) vs. mean: 4.86 (SD: 0.85) log HIV-RNA copies/ml for the T/T] (p = 0.04), and for the HLA-B*5701 (+) [mean: 3.4 (SD: 0.49)] compared to the HLA-B*5701 (-) cases [mean: 4.6 (SD: 0.95)]. In patients without HCV coinfection, HLA-B*5701 (+) genotype was associated with higher baseline CD4+ T cell counts [median: 537 (IQR: 361–682) cells/μl] vs. median 208 (IQR: 61–500) cells/μl for the HLA-B*5701 (-) (p = 0.012), higher CD4+ T cell nadir [median: 437 (IQR: 313–628) cells/μl] vs. median 162 (IQR: 45–386) cells/μl (p = 0.08) and lower pretreatment viral loads (mean: 3.87 [SD: 1.3] vs. mean: 4.97 [SD: 0.91] log HIV-RNA copies/ml (p = 0.004). The remaining associations were not significant when analyzed separately for HIV/HCV coinfected and HIV monoinfected subgroups.2 = 0.321, p<0.0001) than among T/T homozygotes   versus r2 = 0.268, p<0.0001 for HLA-B*5701(-)]   Fig  and 3b, (-)] Fig  and 3d. 019) Fig  and 3f.HCP5 locus favorable for the viremia set-point in Caucasians, was not confirmed for Afro-American population [Favorable effects of certain HLA class I alleles on the outcomes of HIV infection have been shown in many studies , 22, 23.pulation .Moreover, some studies were conducted in the populations infected with divergent HIV subtypes ,25. Our We found that the -35 SNP rs9264942 allele C is the strongest marker for the lower baseline HIV plasma viremia as well as higher CD4+ T cell count and CD4+ T cell nadirs. It is consistent with the previous studies presenting advantageous influence of this polymorphism , 22, 28.The novel finding of this report includes the observation that the HLA-B*5701(+)/HLA-C rs9264942 C/C haplotype is associated with the most favorable pattern of influence on HIV viremia and CD4+ T cell count among the analyzed variants. This haplotype is clearly linked with the lowest pretreatment plasma viral loads, the highest CD4+ T cell counts at care-entry, as well as with the highest nadir of CD4+ T cell count, and reflects delayed progression to immunodeficiency.We found that the coexistence of two protective HLA gene variants results in an additive protective effect on HIV infection. Some studies revealed that HLA-C variants independently control the HIV infection ,13, 22, The protective host genes exert immune pressure on the virus, which creates “escape” mutations. Such finding connected with C/C variant of rs9264942 was presented by Blais and colleagues . The freThe limitation of the study is related to the fact, that it was impossible to assess the time from infection to the care entry, therefore differences in HIV plasma viremia and CD4+ T cell counts related to the time of infection in association with investigated HLA variants were not analyzed.To conclude, we have confirmed the protective effect during the HIV infection of the analyzed HLA variants, including additive one for the compound haplotype. Analysis of genetic variants across various ethnic groups allows confirming the consistence of the genetic effect exerted by the investigated variants.


It has been unclear whether thrombolytic-related asymptomatic hemorrhagic transformation (AHT) affects the clinical outcome. To answer this question, we examined whether thrombolytic-related AHT affect short-term and long-term clinical outcome.All data were collected from the Thrombolysis Implementation and Monitor of Acute Ischemic Stroke in China (TIMS-China) registry. The patients were diagnosed as having AHT group and non- hemorrhagic transformation (HT) group based on clinical and imaging data. The patients with symptomatic hemorrhagic transformation were excluded from this study. Thrombolytic-related AHT was defined according to European-Australasian Acute Stroke Study (ECASS) II criteria. 90-day functional outcome, 7-day National Institutes of Health Stroke Scale (NIHSS) score, 7-day and 90-day mortalities were compared between two groups. Logistic regression analysis was used to evaluate the effects of AHT on a short-term and long-term clinical outcome.904 of all 1440 patients in TIMS-China registry were enrolled. 89 (9.6%) patients presented with AHT after thrombolysis within 24-36h. These patients with AHT were more likely to be elder age, cardioembolic subtype, and to have higher National Institutes of Health Stroke Scale score before thrombolysis than patients without AHT. No significant difference was found on the odds of 7-day (95% CI:0.692 (0.218–2.195), (P = 0.532) or 90-day mortalities (95% CI:0.548 (0.237–1.268), P = 0.160) and modified Rankin Score(0–1) at 90-day (95% CI:0.798 (0.460–1.386), P = 0.423) or modified Rankin Score(0–2) at 90-day (95% CI:0.732 (0.429–1.253), P = 0.116) or modified Rankin Score(5–6) at 90-day (95% CI:0.375 (0.169–1.830), P = 0.116) between two groups.Thrombolytic-related AHT does not deteriorate short-term and long-term clinical outcome. Intravenous thrombolysis with alteplase (recombinant tissue plasminogen activator) is the most effective therapy for acute ischemic stroke ,2. HowevAll of data were collected from the Thrombolysis Implementation and Monitor of acute ischemic Stroke in China (TIMS-China) registry , a natioEligible patients received intravenous thrombolysis within 4.5-hour. Patients with incomplete data and thrombolysis time window>4.5h or unknown were excluded. Patients with NIHSS score ≥25were excluded. Patients with symptomatic HT were excluded from this study. Eligible patients were divided into two groups: the AHT group, which had asymptomatic hemorrhagic transformation; and the non-HT group, which had no thrombolytic-related hemorrhagic transformation .The registry of TIMS-China was approved by ethics committee of Beijing Tiantan hospital in 2006. The data from the TIMS-China registry was anonymous before access and analysis by all researchers and patient treatment was assigned and determined independent of all researchers. All patients had written informed consent before being entered in the registry.Follow-up CT scans for patients were collected at 24–36 hours and 7-day. The diagnosis of HT on brain images was determined independently by 2 neurologists blinded to clinical data. If there was a disagreement between 2 neurologists, a third neurologist was consulted, and a consensus decision was reached. Hemorrhagic transformation must be scored according to the ECASS II classification [Results were reported as mean ± SD or the frequency of categorical variables. Pearson chi-square (χ2) test was used for comparisons of categorical variables. Continuous variables were analyzed using t test or Mann-Whitney U test. Percentage proportions of outcome events were calculated by dividing the number of events by the total number of patients. For each outcome variable, a separate multivariable logistic regression was performed. Odds ratios (ORs) with its 95% confidence intervals (CI) were calculated by using non-HT group after thrombolysis as the reference group. Two-sided values of P less than .05 were considered statistically significant. All statistical analyses were performed using SAS software version 9.3 .904 patients (62.8%) were enrolled from a patient population of 1440 from 67 centers across TIMS-China. Our exclusion criteria were as follows: (1) incomplete neuroimaging workups (n = 40), (2) loss to follow-up (n = 36), (3) thrombolysis time window>4.5h or unknown (n = 312), (4) symptomatic HT (n = 24), (5) Patients with severe stroke with a NIHSS score >25(n = 93). It had been reported that 89(9.6%) patients were transformed to AHT in China , 24  and mRS score (0–2) in patients after thrombolysis when we analyze the data without any adjustment for covariates. After adjustment for baseline relevant factors, no significant difference was found on the odds of 7-day (95% CI:0.692 (0.218–2.195), (P = 0.532) or 90-day mortalities (95% CI:0.548 (0.237–1.268), P = 0.160) and modified Rankin score(0–1) at 90-day (95% CI:0.798 (0.460–1.386), P = 0.423) or modified Rankin score(0–2) at 90-day (95% CI:0.732 (0.429–1.253), P = 0.116) or modified Rankin score(5–6) at 90-day (95% CI:0.375 (0.169–1.830), P = 0.116) between two groups . In AHT There were remarkably low rates of asymptomatic hemorrhagic transformation 4.5%)  in the N.5%  in tPrevious some studies have shown that thrombolytic-related AHT does not have a negative effect on the short-term or long-term functional outcome. AHT after thrombolysis has a suggestive of vascular recanalization and effective treatment . MoreoveDifferent from mechanism of symptomatic HT, we speculate that AHT may link with small artery formation rather than large artery formation. It has been showed that thrombolysis after acute ischemic infarction may lead to artery reopening and collateral circulation reperfusion6 . MoreoveOur study may have the following limitations. (1) Our sample size was relatively small, and our data exclude severe stroke(NIHSS> or = 25), which maybe have a subtle effect (either positive or negative) of AICH outcome. (2) Another limitation is that the study did not follow the randomized, double-blind principles. Despite these limitations, our results have implications that post-thrombolytic AHT in China does have not a negative effect on clinical prognosis.S1 Table(DOCX)Click here for additional data file.S2 Table(XLS)Click here for additional data file.


There is a lack of data on potential gender differences in the use of interventions to prevent and treat cardiovascular disease (CVD) in HIV-positive individuals. We investigated whether such differences exist in the D:A:D study.Follow-up was from 01/02/99 until the earliest of death, 6 months after last visit or 01/02/13. Rates of initiation of lipid-lowering drugs (LLDs), angiotensin-converting enzyme inhibitors (ACEIs), anti-hypertensives and receipt of invasive cardiovascular procedures  were calculated in those without a myocardial infarction (MI) or stroke at baseline, overall and in groups known to be at higher CVD risk: (i) age >50, (ii) total cholesterol >6.2 mmol/l, (iii) triglyceride >2.3 mmol/l, (iv) hypertension, (v) previous MI, (vi) diabetes, or (vii) predicted 10-year CVD risk >10%. Poisson regression was used to assess whether rates of initiation were higher in men than women, after adjustment for these factors.n=13,039; median (interquartile range) 34 (29–40) years) were younger than men  years, p=0.001), and were less likely to be current smokers , to have diabetes  or to have hypertension . Of 49,071 individuals without a MI/stroke at enrolment, 0.6% women vs. 2.1% men experienced a MI while 0.8% vs. 1.3% experienced a stroke. Overall, women received ICPs at a rate of 0.07/100 person-years (PYRS) compared to 0.29/100 PYRS in men. Similarly, the rates of initiation of LLDs (1.28 vs. 2.46), anti-hypertensives (1.11 vs. 1.38) and ACEIs (0.82 vs. 1.37) were all significantly lower in women than men (At enrolment, women (than men . As expethan men . Use of most CVD interventions was lower among women than men in the D:A:D study. Our findings suggest that actions should be taken to ensure that both men and women are monitored for CVD and, if eligible, receive appropriate CVD interventions.


Pressure-support ventilation improves lung mechanics, blood gas exchange, hemodynamics, and work of breathing (WOB) in mild acute respiratory distress syndrome (ARDS) , 2. NeveT=6ml/kg) using two PEEP levels (2 and 5 cmH2O) in a mild ARDS model.To compare PSV and PCV target to protective tidal volume  instillation. After 24 hours, animals were anesthetized, tracheotomized, and their lungs were mechanically ventilated in PSV to achieve VT = 6 ml/kg. After baseline data collection, animals were randomly divided to four groups (n=8/group):2O (PCV-P2);PCV + PEEP = 2 cmH2O (PCV-P5);PCV + PEEP = 5 cmH2O (PSV-P2);PSV + PEEP = 2 cmH2O (PSV-P5).PSV + PEEP = 5 cmHThirty-two male Wistar rats (310 ± 19 g) were submitted to intratracheal RS) and peak transpulmonary  pressures, and pressure-time product (PTP), as a surrogate of WOB, were evaluated.Animals were ventilated for 2 hours. Mean arterial pressure (MAP), arterial blood gases, peak airway . In accordance, PTP was lower in animals submitted to PEEP = 5 cmH2O compared to PEEP = 2 cmH2O during PSV .All animals showed better oxygenation along time, regardless of ventilator strategy. Animals submitted to PCV, regardless of PEEP, received more colloids to keep MAP>70 mmHg. Ppeak,In a mild ARDS model, pressure-support ventilation is associated to better hemodynamics, lung mechanics, and it seems to have a dependent effect of the adjusted PEEP level, as depicted by work of breathing.CNPq, FAPERJ, CAPES, PRONEX, MS-DECIT


Systolic myocardial strain is load dependent, but the CMR literature largely disregards effects of myocardial afterload on strain and strain rate. This may reflect well known limitations of conventional myocardial afterload assessment using wall stress analyses, which are based on erroneous assumptions about left ventricular(LV) geometry and/or myocardial material properties. Therefore, we compared the utility of a nongeometric afterload index(NGI) derived from LV pressure(P) volume(V) and mass, which requires no assumptions about material properties, to that of conventional noninvasive end-systolic circumferential stress(CWS) as determinants of CMR LV circumferential strain(CST), ejection fraction(EF) and strain rate(SR) in normals(NL) and patients with nonischemic dilated cardiomyopathy(CM).We obtained breath-hold volumetric short-axis SSFP cines, cuff systolic P, systolic duration, LVV, M and EF, feature-tracking global CST(TomTec Imaging Systems) and mean SR in NLsyrs) and CM  yrs, EF 27.2%(sd10.8%). CWS was calculated using Mirsky's formula(Biophys. J.1969) while NGI was determined as end-systolic PV/M.EF, CST and CSR were markedly reduced in CM compared to NL(EF 27.2%sd10.8)vs 58.4%(4.6), (-53%)p < 0.0001; CST -10.7%(5.3) vs -23.9%(4.3), (-55%), p < 0.0001);(CSR -32.1%/s(14.8) vs -65.7(14.9) p < 0.0001). But CWS was also markedly elevated in CM versus NL(CWS 307.6(9.2) vs 176.2(42.1)x 103 dyn/cm2,(+75%), p < 0.0001). Thus afterload excess due to adverse LV remodeling, may account for most EF, strain and strain rate reduction in CM. However, PV/M was more markedly increased than CWS, (162.6(sd48.9) vs 84.4(18.4)(+93%) p < 0.0001) and correlated more closely and significantly with EF, CST and CSR than CWS in the expected inverse relationship in both NL and CM subgroups(Table 0.8vs 58.Afterload excess due to adverse LV remodeling is an important determinant of reduced myocardial and LV chamber function in CM, making a major contribution to reductions in EF, CST and SR, but PV/M, a simple, nongeometric afterload index, is superior to conventional wall stress calculation as a quantitative afterload index.St. Francis Research Foundation.


P1 = 0.007; CR vs. SD: P2 < 0.001; PR vs. SD: P3 = 0.004), 95.7% vs. 88.7% vs. 70.2% , 92.0% vs. 87.4% vs. 74.3%  and 95.9% vs. 88.8% vs. 81.8% , respectively. Multivariate analysis identified that the tumour response to IC was an independent prognostic factor for DFS, OS and LRRFS.The prognostic value of the tumour response to induction chemotherapy (IC) for long-term survival outcomes after intensity-modulated radiation therapy in nasopharyngeal carcinoma (NPC) remains unknown. We retrospectively reviewed 1811 consecutive patients with newly diagnosed NPC treated using IMRT, and 399 eligible patients with pre- and post-induction chemotherapy magnetic resonance images were recruited. The clinicopathological features of patients with different tumour responses were compared using the Chi-square test or Fisher’s exact test. Prognostic value was assessed using a multivariate Cox proportional hazards model. After IC, 101/399 (25.3%) patients had a complete tumour response overall (CR), 262 (65.7%) had a partial response (PR) and 36 (9.0%) had stable disease (SD). The 4-year disease-free survival (DFS), overall survival (OS), distant metastasis-free survival (DMFS) and locoregional relapse-free survival (LRRFS) rates for CR vs. PR vs. SD were 90.0% vs. 79.0% vs. 58.2% (CR vs. PR:  Nasopharyngeal carcinoma (NPC) is a cancer with an extremely unbalanced geographical distribution, with an age standardised incidence rate of 20–50 per 100,000 males in south China2345et al. reported that neoadjuvant docetaxel-cisplatin followed by CCRT provided a 3-year overall survival (OS) benefit in stage III-IVB NPC891011The value of additional induction chemotherapy (IC) before CCRT in advanced NPC has received intense investigation. In 2009, Hui et al. revealed that the unsatisfactory tumour response after induction chemotherapy could predict poor prognosis for patients with advanced-stage NPCHowever, despite these negative outcomes, IC may have potential clinical value. Previous studies reported that the response to chemotherapy correlate with clinical outcome1314Of the 1811 patients with newly diagnosed non-metastatic NPC treated between November 2009 and February 2012 at Sun Yat-sen University Cancer Center, and the 399 patients for whom both pre- and post-induction chemotherapy magnetic resonance (MR) images of the nasopharynx and cervical region were available were retrospectively analysed. This study was conducted in compliance with the institutional policy regarding the protection of patients’ private information and approved by the Research Ethics Committee of Sun Yat-sen University Cancer Center. All the methods were carried out in accordance with the approved guidelines of Sun Yat-sen University Cancer Center. Written informed consent was obtained from all patients prior to therapy.Routine staging workup included a complete history, clinical examinations of the head and neck, direct fibre-optic nasopharyngoscopy, magnetic resonance imaging (MRI) of skull base and whole neck, chest radiography, whole-body bone scan, abdominal sonography, and positron emission tomography-CT if clinically-indicated. Immunoglobulin A antibodies against EBV viral caspid antigen (VCA-IgA) and Epstein Barr virus early antigen (EA-IgA) were quantified. All patients underwent dental evaluations before RT.th edition of the International Union against Cancer/American Joint Committee on Cancer (UICC/AJCC) staging systemPatients were restaged according to the 7b-values = 0 and 1000 s/mm2; three signal averages) were obtained before contrast injection. After intravenous administration of gadopentetate dimeglumine , axial and sagittal T1-weighted spin-echo and coronal T1-weighted fat-suppressed spin-echo sequences were performed .All patients underwent MRI of the region from the suprasellar cistern to the inferior margin at the sternal end of the clavicle using a head-and-neck coil with a 3 Tesla system . T1-weighted fast spin-echo images in the axial, coronal and sagittal planes (repetition time [TR]/echo time [TE] = 650 ms/9 ms), T2-weighted fast spin-echo MR images in the axial plane (TR/TE = 2470 ms/90 ms) and a spin-echo echo-planar DWI sequence All patients underwent IMRT while immobilized using a custom head-to-neck thermoplastic cast with the patient’s neck resting on a support. A high-resolution contrast planning CT scan was taken from the vertex to 2 cm below sternoclavicular joint . Target volumes were delineated slice-by-slice on treatment planning CT scans using an individualized delineation protocol that complies with International Commission on Radiation Units and Measurements reports 50 and 62. Prescribed doses were 66–72 Gy at 2.12–2.43 Gy/fraction to planning target volume (PTV) of primary gross tumour volume (GTVnx), 64–70 Gy to PTV of GTV of involved lymph nodes (GTVnd), 60–63 Gy to PTV of high-risk clinical target volume (CTV1), and 54–56 Gy to PTV of low-risk clinical target volume (CTV2). All targets were treated simultaneously using the simultaneous integrated boost technique.2) with 5-fluorouracil (1000 mg/m2) (PF), cisplatin (75 mg/m2) with docetaxel (75 mg/m2) (TP), or cisplatin (60 mg/m2) with 5-fluorouracil (600 mg/m2) and docetaxel (60 mg/m2) (TPF) every three weeks for two or three or more cycles. Concurrent chemotherapy was cisplatin weekly (30–40 mg/m2) or on weeks 1, 4 and 7 (80–100 mg/m2) of radiotherapy.According to institutional guidelines, we recommended RT alone for stage I, concurrent chemoradiotherapy (CCRT) for stage II, and CCRT +/− IC/adjuvant chemotherapy (ACT) for stage III to IVA-B. IC consisted of cisplatin . The end points (time to first defining event) were disease-free survival (DFS), overall survival (OS), distant metastasis-free survival (DMFS), locoregional relapse-free survival (LRRFS), local relapse-free survival (LRFS) and regional relapse-free survival (RRFS).P < 0.05 was considered significant. Stata Statistical Package 12  was used for all analyses.The Chi-square test or Fisher’s exact test were used to compare categorical variables and treatment failure patterns between the CR, PR and SD groups. Life-table estimation was performed using the Kaplan-Meier method and log-rank test. The multivariate Cox proportional hazards model was used to estimate hazard ratios (HRs) and 95% confidence intervals (CIs); age, gender, smoking, drinking, pathological type, T category, N category, concurrent chemotherapy, overall response were included as variables. All tests were two-sided; n = 308)-to-female (n = 91) ratio was 3.4:1. The median age for the whole cohort was 45 years . The baseline characteristics of patients were list in For the 399 patients, the male  patients, PR for 262 (65.7%) patients, and SD for 36 (9.0%) patients. No patients had PD after IC. The local tumour response was a CR for 139/399 (34.8%) patients, PR for 195 (48.9%) patients and SD for 65 (16.3%) patients. Nineteen (4.8%) patients had N0 disease and were not included in the analysis of regional response. With regards to regional tumour response, 167/380 (43.9%), 166/380 (43.7%) and 47380 (12.4%) patients had a CR, PR and SD, respectively.P = 0.002; 58.3% vs. 40.1%, P = 0.038) and had advanced T category  compared to patients who achieved a CR or PR. Obviously, patients with CR received more cycles of induction chemotherapy compared with that of patients with PR (P = 0.009) or SD (P < 0.001). No significant associations were observed between any other clinicopathological feature and the overall tumour response.Significantly more patients with SD were previous smokers . By final follow-up, 21/399 (5.3%) patients had developed local failure, 23/399 (5.8%) patients developed regional failure and 51/399 (12.8%) patients developed distant failure.P = 0.022) and distant failure  after IMRT than patients with a CR (P = 0.054).With regards to overall tumour response after induction chemotherapy, patients with SD had higher rates of local failure . The local failure rates of the local PR and CR groups were almost significantly different . Patients with a regional CR after induction chemotherapy had a lower rate of distant failure after IMRT than patients with a regional PR  and regional SD . Additionally, patients with a regional CR had a lower rate of regional failure after IMRT than patients with a regional PR .Patients with local SD after induction chemotherapy had higher rates of local failure after IMRT compared to patients with a local CR , 95.7% vs. 88.7% vs. 70.2% , 92.0% vs. 87.4% vs. 74.3%  and 95.9% vs. 88.8% vs. 81.8% , 92.3% vs. 89.2% vs. 80.6% , 97.7% vs. 94.2% vs. 88.5%  and 90.5% vs. 85.7% vs. 85.9% , 93.7% vs. 88.7% vs. 71.5% , 97.5% vs. 91.1% vs. 93.1%  and 92.6% vs. 86.4% vs. 73.5%  is associated with a poorer prognosis in NPC, breast cancer and gastric cancerNPC is highly chemosensitive and radiosensitive. Although most patients with advanced NPC respond well to chemotherapy, recurrence of distant metastases is the major cause of treatment failure and has a poor prognosisDespite the outcome that unsatisfactory (PR or SD) tumour response after IC indicated poor prognosis, we should pay attention to the factors which may affect the result. Firstly, using the RECIST standardThe current study showed that patients with unsatisfactory tumour responses after IC have poorer prognosis compared with that of patients with satisfactory tumour responses. Therefore, we should pay more attention to patients with SD after induction chemotherapy in clinical practice. More intensive treatment regimens like higher radiation dose and adjuvant chemotherapy should be considered. Moreover, regular follow-up should also be provided to detect early recurrence and reduce the potential risk of distant failure.This study is limited by its retrospective nature and fact the follow-up time may have been insufficient, though we selected DFS as the major endpoint to address this shortcoming. Moreover, many other prognostic factors including pre-treatment plasma Epstein-Barr virus DNA load272830Tumour response to induction chemotherapy is an independent prognostic factor for patients with NPC receiving IMRT. Assessing tumour response after induction chemotherapy may assist prognostication and refine the treatment of high-risk patients with NPC. The outcomes in this current study need to be confirmed by prospective studies with large cohorts.How to cite this article: Peng, H. et al. The Tumour Response to Induction Chemotherapy has Prognostic Value for Long-Term Survival Outcomes after Intensity-Modulated Radiation Therapy in Nasopharyngeal Carcinoma. Sci. Rep.6, 24835; doi: 10.1038/srep24835 (2016).


Mucuna pruriens is the best known natural source of L-dopa, the gold standard for treatment of Parkinsonism. M. pruriens varieties are protein rich supplements, and are used as food and fodder worldwide. Here, we report L-dopa contents in seeds of fifty six accessions of four M. pruriens varieties, M. pruriens var. pruriens, M. pruriens var. hirsuta, M. pruriens var. utilis and M. pruriens var. thekkadiensis, quantified by HPTLC-densitometry. L-dopa contents varied between 0.58 to 6.42 . High and low L-dopa yielding genotypes/chemotypes of M. pruriens could be multiplied for medicinal and nutritional purposes, respectively. HPTLC profiles of M. pruriens seeds on repeated extraction (24 h) in 1:1 formic acid-alcohol followed by development in butanol:acetic acid:water  showed consistent degradation of L-dopa (Rf 0.34 ± 0.02) into a second peak (Rf 0.41 ± 0.02). An average of 52.11% degradation of L-dopa was found in seeds of M. pruriens varieties. Since M. pruriens seeds and/or L-dopa are used for treatment of Parkinson’s disease and as an aphrodisiac both in modern and/or traditional systems of medicine, the finding of high level of L-dopa degradation (in pure form and in M. pruriens extracts) into damaging quinones and ROS is very significant. Mucuna pruriens (L.) DC. is a climbing legume distributed across the tropics. Four varieties of the species have been documented so far from south India, of which M. pruriens var. pruriens is well distributed1M. pruriens var. hirsuta and M. pruriens var. thekkadiensis are restricted to the southern parts of the Indian peninsula, and M. pruriens var. utilis occurs only in cultivationM. pruriens var. pruriens and Mucuna pruriens var. utilis find importance as food, feed, cover crop and fodder and are extensively cultivated worldwide345M. pruriens varieties propagate mostly through their seeds. M. pruriens var. pruriens is best known as the natural source of the aromatic amino acid, L-3,4-dihydroxy phenylalanine (levodopa or L-dopa)  . L-dopa L-dopa and dopamine, generating semiquinones, quinones, oxygen radicals and other reactive oxygen species (ROS), play a role in neuronal cell death in PD20O-quinone products of L-dopa autoxidation are cytotoxic to cellular systems222324M. pruriens var. pruriens and L-dopa could recover spermatogenic loss which makes them the treatment of choice for infertility27PD is characterized by degeneration of dopaminergic neurons in the substantia nigra, and subsequent deficiency of the neurotransmitter dopamine in the brain areas. PD affects motor activities including writing and speaking abilities. Recent studies suggested oxidative stress, mitochondrial dysfunction and impairment of the ubiquitin-proteasome system as the major factors involved in pathogenesis of PD7891011et al., 2011 and Sundaram and Gurumoorthi, 2012 standardized protocols for HPTLC-based quantification of L-dopa in M. pruriens var. pruriens seeds29et al., 2008 , Behara et al., 2010 (4.83%), Raina and Khatri, 2011 (2.23–5.36%) and Raina et al., 2012 (3.29–5.44%) quantified L-dopa contents in M. pruriens var. pruriens seeds by HPTLC313233et al., 2007et al., 2011et al., 2008et al., 2008et al., 2010M. pruriens var. pruriens seeds (7.20%) and its formulations (4.20–5.60%) by spectroflourimetryStizolobium pruriens var. utilis (M. pruriens var. utilis) (3.9–10.6%)M. pruriens var. utilis (4.39–5.21%)M. pruriens var. pruriens (4.0–6.0%)et al., 2012 quantified L-dopa in M. pruriens var. pruriens formulations (3.0–6.0%) by HPLCet al., 2010 developed HPLC-based quantification of L-dopa in M. pruriens var. utiliset al., 2010 carried out HPLC-MS/MS quantification of L-dopa in rat plasmaet al., 1997 determined threshold L-dopa levels in plasma of patients with advanced PD by in vitro microdialysis-HPLCVachhani 1 2.23–5.% and Raiutilis 4.–5.21%M. M. pruriens seeds and formulations by chromatographic techniques  are on limited number of samples and involved prolonged, multistep extraction procedures in acidic media. Screening of more M. pruriens varieties/accessions could lead to the discovery of high (elite) and low L-dopa yielding accessions suitable for medicinal and nutritional purposes, respectively. Secondly, in our preliminary HPTLC profiling of M. pruriens extracts and L-dopa standard, we repeatedly detected labile L-dopa-based degradation signals. Most similar HPTLC/HPLC studies never recorded this degradation signal of L-dopa562931323334353638394041M. pruriens varieties viz., M. pruriens var. pruriens (21), M. pruriens var. hirsuta (3), M. pruriens var. utilis (5) and M. pruriens var. thekkadiensis (1), collected from various locations in Kerala in south India, (ii) L-dopa contents in second generation seeds of twenty-six accessions of M. pruriens var. pruriens (21) and M. pruriens var. utilis (5) grown in an Experimental Plot (EP) under identical ecological conditions, (iii) degradation patterns of L-dopa in M. pruriens seed extracts, (iv) quantification of L-dopa degraded products in seeds  of four M. pruriens varieties and (v) characterization of L-dopa degraded moieties by HPTLC, DART-MS and LC/EI-MS.Most L-dopa quantification studies in M. pruriens var. pruriens accessions varied from 0.89 to 6.42   to 3.34%. L-dopa contents in wild accessions of M. pruriens var. hirsuta ranged from 1.01 to 4.27%, and % SDP ranged from 0.01 to 1.40%. L-dopa contents in seeds of M. pruriens var. utilis wild accessions varied from 0.65 to 2.67%, and SDP contents varied from 0.12 to 3.82%. In EP grown accessions of M. pruriens var. utilis, L-dopa contents ranged from 1.33 to 3.97%, and SDP contents were zero to 0.77%. M. pruriens var. thekkadiensis wild accession showed 4.34% of L-dopa with SDP 0.01% (M. pruriens var. hirsuta and M. pruriens var. thekkadiensis in the Field Gene Bank (FGB). This is the first report of L-dopa quantification in M. pruriens var. hirsuta and M. pruriens var. thekkadiensis. This is also the first quantitative determination of degradation products of L-dopa in M. pruriens seeds.L-dopa contents in wild dr. wt.) . Percentdr. wt.) . In secoDP 0.01% . FruitinM. pruriens var. pruriens, M. pruriens var. hirsuta and M. pruriens var. utilis seeds, L-dopa contents did not show any positive correlation with altitudes of their collection locations   accessions , M. pruriens var. thekkadiensis  and M. pruriens var. hirsuta . Lowest L-dopa percentages were found in EP grown M. pruriens var. pruriens seeds   showed lowest degradation levels . Similarly, lowest L-dopa contents  were seen in seeds with high levels of degradation  (In wild %/2.96%) . M. prurcessions . Highest, 0.84%) . Again, , 2.75%) .M. pruriens accessions was 52.11% of total L-dopa content (138.45%)  are highly significant. Standard L-dopa also showed similar degradation into a second signal on HPTLC profile . Previous studies rarely mentionedM. pruriens seeds and formulations2931323334353638394041Total SDP 72.15%) in wild 30) and EP grown 26) 138.45%) . These d and EP g% in wild 138.45%)f 0.34 ± 0.02 (f 0.34 ± 0.02 (L-dopa) and a SDP at Rf 0.41 ± 0.02  showed two significant signals  at varying ratios  M. pruriens var. pruriens seed extract and decomposed L-dopa standard showed degradation patterns on HPTLC, DART-MS and LC/EI-MS  showed M+H+ signals at 123.06 (medium), 154.10 , 162.07 (medium), 198.09 , 199.09 (minor) and 224.11 (minor)  showed M+H+ signals at 129.08 (medium), 176.08 (major), 190.10 (major), 191.11 (minor), 192.13 (minor), 193.13 , 195.17 , 204.11 (major), 218.13 (medium), 244.12 (medium) and 245.13 (minor)  on HPTLC showed only one signal at R4 ± 0.02 . But, L-1 ± 0.02 . M. prurg ratios . Fresh (LC/EI-MS . On DARTt 198.09 . Fresh M (minor) . Decompo (minor) .M. pruriens var. pruriens extract (24 h extracted) showed only major signals of L-dopa and dopamine. Decomposed M. pruriens var. pruriens extract did not show L-dopa and dopamine, instead showed a group of degradation signals at the M+H+ 190.10, 191.11, 192.13, 193.13 and 195.17. These DART-MS signals correspond to dopachrome, leucodopachrome, dopaquinone, other quinones and ROS , 20 mM Tris buffer  and water at various time periods  were tested through HPTLC profiling. Time dependent degradation was observed in M. pruriens var. pruriens extracts and in standard L-dopa at these pH values , M. pruriens var. utilis (10), M. pruriens var. hirsuta (3) and M. pruriens var. thekkadiensis (1) accessions varied between 0.58 to 6.42 . M. pruriens var. pruriens elite accessions are 4283  and 4498   and M. pruriens var. hirsuta  (M. pruriens var. pruriens (4098) showed highest L-dopa content of 4.32%  and M. pruriens var. utilis  accessions, which showed less than 1% L-dopa, can be multiplied and utilized as protein-rich diets.Our study led to the discovery of elite genotypes/chemotypes in 56 accessions of four %, wild) . Other e, 4.27%) . Among tof 4.32% . These eM. pruriens seeds in the acidic medium of 1:1 formic acid-alcohol. On HPTLC profiling, seed extracts of all four M. pruriens varieties (collected from various wild locations and EP grown) showed L-dopa at Rf 0.34 ± 0.02 and a consistent second degradation peak at Rf 0.41 ± 0.02. This degraded peak was indentified as a labile mix of dopamine, dopachrome, leucodopachrome, dopaquinone and other ROS by HPTLC, DART-MS and LC/EI-MS. On an average, the degradation of L-dopa in a 24 h extraction protocol was a high 52.11%, which is significant enough to cause adverse effects in biological systems.We consistently found degradation patterns of L-dopa in M. pruriens var. pruriens seed extract (4450) and L-dopa standard in acidic, neutral and water media showed time dependent degradation. L-dopa degradation rates in the acidic medium  were equivalent or even higher compared to Tris buffer  or water under identical extraction periods. In seven days, M. pruriens var. pruriens seed extract and L-dopa in these liquid media resulted in degradation with gradual appearance of a black deposit. In 30 days, both M. pruriens var. pruriens seed extract and L-dopa in these solvent media resulted in significant degradation and strong black deposits. The degradation rates of L-dopa in M. pruriens var. pruriens extracts and standard L-dopa were low at 4 °C compared to room temperature.HPTLC profiles of M. pruriens seeds or L-dopa are administered for very long periods. Chronic L-dopa therapy in PD results in movement disorders or dyskinesia in most patients. M. pruriens seeds and its preparations are also used for the treatment of PD in the traditional medicinal system of Ayurveda since ancient times. Certain studies claimed that M. pruriens seeds are even more effective than L-dopa in PD treatment. Some literature reports caution insufficient evidence to recommend the clinical use of M. pruriens in the treatment of PD45M. pruriens seeds are also used in tonics for male vitality and virility in Ayurveda28Recent studies showed that L-dopa degradation products and dopamine adducts result in oxidative stress and cause selective cytotoxicity of neuronal cells inducing pathogenesis in PD1820212223242543M. pruriens seeds and/or L-dopa are used for treatment of PD and as an aphrodisiac both in modern and/or traditional systems of medicine, the finding of high level of L-dopa degradation (in its pure form and in M. pruriens extracts) into damaging quinones and ROS is very significant. Our finding of consistent degradation products suggests the need for careful review of the processing of M. pruriens seeds , L-dopa and their mode(s) of administration . Further studies are required to confirm the adverse effects of the degradation products from the ‘cure’  itself in PD patients and other users.Since M. pruriens varieties viz. M. pruriens var. pruriens (21), M. pruriens var. hirsuta (3), M. pruriens var. utilis (5) and M. pruriens var. thekkadiensis (1) were collected in January to April 2009 from various wild locations in Kerala in south India (M. pruriens accessions were recorded during field trips. Voucher specimens of these M. pruriens accessions were deposited at the Herbarium (TBGT) of Jawaharlal Nehru Tropical Botanic Garden and Research Institute (JNTBGRI). Seeds of these thirty M. pruriens accessions collected were dried and powdered (separately).Seeds of thirty accessions of th India . GPS cooM. pruriens seeds collected were initially planted in a FGB of the species established at JNTBGRI. First generation seeds of 21 accessions of M. pruriens var. pruriens and 5 accessions of M. pruriens var. utilis were collected from the FGB and planted in the EP in Randomized Block Design with two replications of each accession and one plant in each replication. M. pruriens var. hirsuta and M. pruriens var. thekkadiensis accessions did not produce fruits in the FGB. Therefore, these two varieties were not planted in the EP. M. pruriens var. pruriens and M. pruriens var. utilis accessions were maintained in the EP in uniform conditions. M. pruriens accessions planted in both FGB and EP were irrigated as and when required, and not supplemented with any external fertilizers. Second generation seeds of 21 accessions of M. pruriens var. pruriens and 5 accessions of M. pruriens var. utilis were collected in January to April 2011, dried and powdered (separately) (A second set of arately) .M. pruriens seed powder (2 g each) was extracted with 1:1 formic acid-alcohol  at room temperature and the extract was filtered. Seed powder residue was then repeatedly extracted with 1:1 formic acid-alcohol , and extracts were filtered. This seed residue was again extracted with 1:1 formic acid-alcohol . Filtrates (of five extractions) were pooled, centrifuged  and made up to 100 ml using 1:1 formic acid-alcohol. This M. pruriens seed extract (5 ml) was concentrated on a rotary evaporator, and the extract weight was recorded. This (concentrated) M. pruriens seed extract was dissolved in 20 ml 1:1 formic acid-alcohol and used for L-dopa quantification by HPTLC-densitometry. This extraction protocol was followed for quantification of L-dopa in seeds of all (56) M. pruriens accessions  made up of Linomat V sample applicator, twin-trough plate development chamber, TLC Scanner 3 and WinCATS Software 4.03. M. pruriens seed extract (5 ml concentrated seed extract) was dissolved in 20 ml of 1:1 formic acid-alcohol (see L-dopa extraction), 4 μl of this solution was repeatedly applied to silica gel HPTLC plate  as 6 mm wide bands with Camag Linomat V sample applicator, fitted with a microsyringe, in N2 flow . L-dopa standard was also applied along with M. pruriens seed extracts. HPTLC plate was developed upto 80 mm in the twin-trough glass chamber pre-saturated for 30 min with mobile phase butanol:acetic acid:water . Developed plate was scanned densitometrically at 282 nm (deuterium lamp) using TLC Scanner 3 equipped with WinCATS software. L-dopa at Rf 0.34 ± 0.02 (n = 56) and a second degradation peak (SDP) at Rf 0.41 ± 0.02 (n = 56) were found in M. pruriens seed extract in butanol:acetic acid:water   it showed a clear second signal at Rf 0.41  . Similar Rf 0.41 . Other s signals . Solventf values  of the standard was reproducible, and was found to be same as the values observed for the peak (L-dopa) in M. pruriens seed extracts. Calibration curve was plotted between amount of standard L-dopa (fresh) versus average response (peak area) . Linearity of the calibration curve in the range 100-1000 ng was ensured. Percentage L-dopa content(s)  in M. pruriens extracts were calculated from peak areas using the standard curve. Percentage of second degradation peak  which is a combination of labile molecules was also quantified based on L-dopa standard curve. Repeatability of sample application  was assessed by applying a sample solution  on a HPTLC plate developed up to 80 mm under saturation conditions with butanol:acetic acid:water  as the mobile phase in the twin-trough glass chamber (previously saturated with the solvent for 30 min). The spot (L-dopa) was scanned six times, % coefficient of variation was acceptable. Robustness of the method was checked by slightly altering the mobile phase composition and plate developing distance was checked. No considerable effect on the data was found. Recovery studies were carried out (in two modes) by the addition of L-dopa to pre-analyzed M. pruriens extracts and they were again analyzed (see Quantification of L-dopa). In the first mode, (i) M. pruriens var. pruriens seed powder  was extracted in the five-step protocol (24 h) and (ii) standard L-dopa (10 mg) was added initially to M. pruriens var. pruriens seed powder  and extracted in the five-step protocol (24 h). (i), (ii) and (iii) fresh standard L-dopa (1 μg/μl), in 1:1 formic acid-alcohol were loaded (4 μl each) onto HPTLC plate, developed with butanol:acetic acid:water  and peak areas were measured at 282 nm . % recovery of L-dopa was calculated from peak areas as 49.78%. In the second mode, M. pruriens var. pruriens seed powder  was extracted in the standard five-step protocol (24 h). (i) M. pruriens var. pruriens extract in 1:1 formic acid-alcohol (4 μl), (ii) M. pruriens var. pruriens extract in 1:1 formic acid-alcohol (4 μl) and fresh L-dopa dissolved in 1:1 formic acid-alcohol  and (iii) fresh L-dopa dissolved in 1:1 formic acid-alcohol  were loaded onto HPTLC plate ((ii) co-spotted), developed and peak areas were measured. % L-dopa recovery was calculated as 99.30%. % residual standard deviations (RSD) were determined as 2.63 (L-dopa) and 3.58 (SDP). Limit of detection  and limit of quantification  were determined for both L-dopa  and SDP 48HPTLC-based quantification of L-dopa was validated in terms of precision, accuracy, repeatability and linearity. Specificity of the assays was tested by repeated application of standard L-dopa. RM. pruriens var. pruriens seed extract (Acc. No. 4450) prepared by the five stage extraction for 24 h (fresh M. pruriens var. pruriens seed extract) and standard L-dopa (fresh) were suspended in 1:1 formic acid-alcohol and kept at room temperature for seven days with occasional stirring. These resulted in decomposed M. pruriens var. pruriens seed extract and decomposed L-dopa standard, respectively. Fresh M. pruriens var. pruriens seed extract, freshly prepared L-dopa standard , decomposed M. pruriens var. pruriens seed extract and decomposed L-dopa standard were applied onto silica gel plates  by HPTLC , developed in butanol:acetic acid:water  and scanned at 282 nm  (zerland) .M. pruriens var. pruriens seed extract (100 mg), decomposed M. pruriens var. pruriens seed extract (100 mg) and decomposed L-dopa standard (26.1 mg) were analyzed by DART-MS on an AccuTOF JMS-T100LC Mass Spectrometer having a DART . Samples were analyzed directly in front of the DART source. Dry He was used at a flow rate of 4 LPM for ionization at 350 °C. Orifice 1 was set at 28 V, spectra were collected, and the data from 6-8 scans were averaged (M. pruriens var. pruriens seed extract (80 mg) and decomposed L-dopa standard (20 mg) were subjected to LC/ESI-MS analysis on a Surveyor-LCQ Deca XP plus system  with Hypersil BDS C18 column , mobile phase: methanol-water 85:15, inj. vol.: 10 μl, flow rate: 0.3 ml/min, run time: 30 min, LC detection: PDA/UV detector, 280 nm and mass detection: electro spray ionization  seeds (1 g each) were separately extracted with 20 ml (each) of 1:1 formic acid-alcohol (strongly acidic), 20 mM Tris-HCl, 20 mM KCl  and water at room temperature and at 4 °C. Similarly, standard L-dopa (5 mg) was extracted (dissolved) in 10 ml each of these three solvents at room temperature and at 4 °C. These M. pruriens var. pruriens seed/L-dopa extracts (3 μl each) were profiled using HPTLC-densitometry (as described in Quantification of L-dopa) at various time periods viz., 1 h after initiation of extraction, 1, 7 and 30 days after initiation of extraction .


The purpose of this study was to determine if consumption of a powdered form of tart cherries derived from tart cherry skins . The lifters ingested the supplements one time daily (480 mg/d) for 10-d: 7-d pre-exercise, day of exercise, and 48-hr post-exercise. Subjects performed 10 sets of 10 repetitions at 70% of 1RM back squat exercises with 3-min recovery between sets, maintaining equivalent total work performed (p = 0.80) and average daily caloric consumption (p = 0.61) between groups. Isokinetic knee extension/flexion maximal voluntary contractions (MVCs) and fasting blood samples were taken pre-squat workout, 60-min following squat workout as well as after 24-h and 48-h of recovery and analyzed by MANOVA with repeated measures.23 resistance trained men  were matched based on relative maximal back squat strength, age, body weight, and fat free mass. Subjects were randomly assigned to ingest in a double blind manner capsules containing a placebo  or powdered tart cherries [CherryPUREPowdered tart cherry supplementation seemed to attenuate the drop from pre-lift measures in 3-repetition summation of isokinetic flexion work , extension work , and all work  through 60-min and 24-h of recovery compared to placebo as reported above by Cohen's d effect size, despite not being statistically significant. The overall MANOVA analysis revealed a significant Wilks' Lambda time (p < 0.001) interaction for all inflammatory markers, but no significant group × time pro-inflammatory (p = 0.30) and anti-inflammatory (p = 0.45) effects. Univariate measures for pro-inflammatory markers reported significant main time effects for TNF-α (p = 0.001), IL-1β (p = 0.30), IL-6 (p = 0.023), and IL-8 (p = 0.018). Univariate measures for anti-inflammatory markers reported significant main time effects for IL-4 (p = 0.001) and IL-7 (p = 0.033) with IL-13 trending toward significance (p = 0.055). No significant group × time effects were observed for any of the inflammatory markers, NT, or TBARS. Serum IL-1β levels were significantly lower in TC compared to P (p = 0.048). Delta changes were assessed at all three recovery time points from the pre-lift marker measures. The overall delta MANOVA analysis revealed a significant Wilks' Lambda pro-inflammatory interaction across time (p = 0.001) and an anti-inflammatory time interaction trending toward significance (p = 0.070), but no significant group × time pro-inflammatory (p = 0.44) or anti-inflammatory (p = 0.30) effects. TNF-α (p = 0.010), IFN-γ (p = 0.042), IL-1β (p = 0.031), IL-6 (p = 0.001), IL-8 (p = 0.025), IL-10 (p = 0.019), and NT (p = 0.018) demonstrated significant main effects on time, while IL-7 approached significance across time (p = 0.095). Serum IL-2 TC (p = 0.074) and IL-10 (p = 0.10) changes from pre-lift tended to be greater across the recovery time coupled with a tendency for IL-10 (p = 0.063) TC levels to also be greater compared to P. Contrarily, serum IFN-γ (p = 0.021) TC changes from pre-lift values were significantly smaller compared to P with specific differences at 24-h and 48-h post-lift.In accordance with previous TC juice supplementation research, the isokinetic performance results of this study indicate that short-term powdered TC consumption 7 days prior to, day of, and 2 days after a single bout of intense resistance exercise may help to attenuate the strength decrement over a 48-h recovery period. The seemingly better maintenance of strength during recovery with short-term powdered TC supplementation surrounding a single bout of resistance exercise did not, however, coincide with any definitive effect on markers of oxidative damage or inflammation. This may be due to the differences in resistance exercise metabolic demands, thus indicating the need for further mechanistic research.


AbstractHoplolaimus Daday, 1905 belongs to the subfamily Hoplolaimine Filipiev, 1934 of family Hoplolaimidae Filipiev, 1934  with ten species, is characterized by lateral field distinct, with four incisures, excretory pore behind hemizonid; Hoplolaimus (Basirolaimus) with 18 species, is characterized by lateral field with one to three incisures, obliterated, excretory pore anterior to hemizonid, dorsal oesophageal gland quadrinucleate; and Hoplolaimus (Ethiolaimus) with four species is characterized by lateral field with one to three incisures, obliterated; excretory pore anterior to hemizonid, dorsal oesophageal gland uninucleate (Hoplolaimuspuriensis Ali, Shaheen & Pervez, 2009 has been described (Hoplolaimus reported in Vietnam, viz H.seinhorsti and H.chambus (The genus ev, 1934 . Daday e in 1905 . Hoplolaal crops . In 199ay, 1905 . Siddiqinucleate . Since tescribed . Up to nchambus .Hoplolaimusbachlongviensis sp. n. was isolated from banana soil in Bach Long Vi Island, Vietnam. The female of this species is described and illustrated below. Some diagnostic characters of this species include body slightly curved ventrally, offset lip region exhibiting three to four annules, lateral field reduced, pharyngeal glands with six nuclei, excretory pore anterior to hemizonid, epiptygma absent, intestine not overlapping rectum and male was not found. Hoplolaimusseinhorsti Luc, 1958 and H.chambus Jairajpuri & Baqri, 1973 were recorded . Soil nematodes were extracted using the decanting and modified Baermann tray method ; IEBR.Nema4050-2 .Hoplolaimusbachlongviensis sp. n. is similar to Hoplolaimusseinhorsti, H.chambus, H.columbus Sher, 1963 and H.pararobustus  Sher, 1963 by having excretory pore anterior to hemizonid, lateral field reduced, represented by interruptions of annules as a single incisure, often indistinct, pharyngeal glands with six nuclei (H.bachlongviensis sp. n. differs from H.seinhorsti by epiptygma absent vs present and number of longitudinal striations on basal ring 6 vs 8-12. It differs from H.chambus by male absent vs present, epiptygma absent vs present, intestine not overlapping rectum vs overlapping rectum. Hoplolaimusbachlongviensis sp. n. differs from H.columbus in having fewer tail annuli 9-13 vs 16-22; a=22-27 vs a=30-38; b=6-8 vs b=9.1-12.4; DGO=3-6 vs DGO=9-13; epiptygma absent vs present; hemizonid located about 7 annuli behind excretory pore vs 2-5 annuli and intestine not overlapping rectum vs overlapping rectum. Hoplolaimusbachlongviensis sp. n. differs from H.pararobustus by male absent vs present, intestine not overlapping rectum vs overlapping rectum, epiptygma absent vs present and sperm absent vs present.x nuclei . HoweverHoplolaimusbachlongviensis sp. n. is distinguished from H.sherivs two incisures in lateral field; having longer stylet 44-50 vs 40-45; having fewer longitudinal striations on basal ring 6 vs 20; hemizonid is conspicuous vs obscure; a=22-27 vs a=26-30; b=6-8 vs b=9.7-11.5.Hoplolaimusbachlongviensis sp. n. differs from H.puriensis by lateral field reduced, represented by a single incisure on the body, but often indistinct vs four lateral lines, longer stylet 44-50 µm vs shorter stylet 32-35 µm.


Enterobacter cloacae strain S1:CND1 isolated from oil-contaminated soil in Guwahati, India. S1:CND1 contains 4,205 coding sequences and has a G+C content of 57.45%. This is the first report of the genome sequence of an E. cloacae adapted to an oil-contaminated environment.We report here the 4.57-Mb draft genome sequence of hydrocarbon-degrading  Enterobacter cloacae is a Gram-negative, rod-shaped bacterium generally associated with nosocomial infections, and is considered one of the most difficult to treat among the Enterobacter sp. , along with 16 rRNAs, 77 tRNAs, 6 noncoding RNAs (ncRNAs), and 50 pseudogenes. Rapid functional annotation for CDSs of strain S1:CND1 was carried out with the RAST annotation server (s = 650 CDSs); amino acids and derivatives (s = 467); stress response (s = 157); respiration (s = 149); fatty acids, lipids, and isoprenoids (s = 138); DNA metabolism (s = 117); regulation and cell signaling (s = 150); protein metabolism (s = 168); RNA metabolism (s = 150); membrane transport (s = 177); virulence, disease, and defense (s = 100); cell wall and capsule (s = 196); and cofactors, vitamins, prosthetic groups, and pigments (s = 253). Genome annotation revealed the presence of hydrocarbon degradation genes as alkane-1-monooxygenase, alkanesufonate monooxygenase, naphthalene 1,2-dioxygenase, and quercetin 2,3-dioxygenase, thus underlining the extensive genetic adaptation of strain S1:CND1 to oil contamination.The genomic DNA for strain S1:CND1 was extracted using an Ultra-Clean Microbial DNA Isolation Kit  according to the manufacturer’s protocol. Isolated genomic DNA was then sequenced with an Illumina HiSeq 2500, which generated 4,580,054 paired-end reads. After quality control measures, the reads were assembled using the  v. 3.81 , SPAdes  v. 3.81 , and Vel v. 3.81 , which gEscherichia coli 88.1467 (score = 501) as its closest neighbor, followed by E. coli 88.0221 (score = 490) and E. coli 89.0511 (score = 469). Enterobacter mori LMG 25706 (score = 344) was identified as the 18th-closest neighbor.A comparison of strain S1:CND1 with genomes in the RAST database identified E. cloacae strain S1:CND1 has been deposited in DDBJ/EMBL/GenBank under the accession no. LUGN00000000. The version of the whole-genome sequence (WGS) described here is version LUGN01000000.This whole-genome shotgun sequencing project for


Thus, AvCystatin ameliorates enhanced RSV pathology without increasing susceptibility to, or persistence of, viral infection and warrants further investigation as a possible therapy for virus-induced airway disease.Respiratory Syncytial Virus (RSV) is a major pathogen causing low respiratory tract disease (bronchiolitis), primarily in infants. Helminthic infections may alter host immune responses to both helminths and to unrelated immune triggers. For example, we have previously shown that filarial cystatin (AvCystatin/Av17) ameliorates allergic airway inflammation. However, helminthic immunomodulators have so far not been tested in virus-induced disease. We now report that AvCystatin prevents Th2-based immunopathology in vaccine-enhanced RSV lung inflammation, a murine model for bronchiolitis. AvCystatin ablated eosinophil influx, reducing both weight loss and neutrophil recruitment without impairing anti-viral immune responses. AvCystatin also protected mice from excessive inflammation following primary RSV infection, significantly reducing neutrophil influx and cytokine production in the airways. Interestingly, we found that AvCystatin induced an influx of CD4 However, due to the proposed antiviral function of neutrophils, current practice suggests to target neutrophils under the umbrella of antiviral treatment .+ T cell derived IL-10. Collectively, our findings support the testing of AvCystatin as a potential novel agent to counter virus-enhanced respiratory inflammation.This is, to our knowledge, the first time that a recombinant parasite-derived immunomodulator has been shown to down-regulate virally-enhanced inflammation without impairing responses important for anti-viral immunity. We provide evidence that AvCystatin immunomodulation may act through the production of CD4All mouse experiments were ethically approved by the Imperial College Central Biological Services (CBS) ethics committee performed in accordance with approved UK Home Office guidelines (Project License No. PPL 70/6785).Mice were treated with 20 μg AvCystatin prepared as previously described  or PBS i5 PFU RSV 2 weeks thereafter. For rvv-G RSV models, mice were primed with vaccinia expressing RSV-G protein (3 x 106 PFU) by scarification of the rump and infected intranasally with 5 x 105 PFU RSV 14 days later. RSV titres were assessed by titration of lung homogenates on Hep-2 cell monolayers [RSV (strain A2) and rVV-G (vaccinia virus expressing the RSV G-protein) were propagated in Hep-2 cells (ATCC) as described previously . FI RSV nolayers . RSV L-gnolayers .BAL-accessible cells were obtained by repeated instillation of 1 ml of PBS/12 mM lidocaine via the trachea in sacrificed animals. Cytospin preparations were stained with hematoxylin and eosin and analysed by light microscopy (300 cells per slide). Lungs were homogenized and digested with collagenase D  for 30 min. Mediastinal  lymph nodes were dissociated and filtered through a 100 μm cell strainer.Flow cytometry was performed as previously described . Briefly6 PEC and infected with RSV i.n. 4 h later.Donor BALB/c were injected i.p. with 20 μg active or heat-inactivated AvCystatin and peritoneal exudate cells (PEC) recovered 20 h later in PBS/2 mM EDTA/0.2% BSA. Recipient mice were injected intravenously with 3x10Quantification of BAL chemokines and cytokines was performed using a 13-plex Luminex kit . Data were acquired with a Luminex 100  and Starstation software.P values of <0.05 were considered significant.GraphPad Prism software  was used to analyse data, generally shown as mean ±SEM of 5 animals per group. Mann-Whitney t-test and 2 way ANOVA were used to compare data. S1 FigP values reflect Mann-Whitney t-test: * p<0.05, **p<0.01.A) Schematic of the FI RSV model: i.m. intramuscular; i.p. intraperitoneal; i.n. intranasal application. B) Weight loss in the FI-RSV model. Total cell number of eosinophil (C) and macrophages in the BAL (D). Viral load in the lungs measured by RSV L-gene copies (E). Representative data of 2 experiments, 5 mice per group. Error bars indicate SEM. (TIF)Click here for additional data file.S2 Fig+ T cells in the BAL (D and E) and lungs (B and C). Representative data of 2 experiments, 5 mice per group. Error bars indicate SEM. P values reflect Mann-Whitney t-test: * p<0.05, **p<0.01.Flowcytometric analysis of IL-10 intracellular cytokine content in RSV challenged or AvCystatin/RSV challenged mice is shown (A). Graphical visualization of IFNγ and IL-10 production by CD4(TIF)Click here for additional data file.S3 Fig+ CD4+ T cells (A) and the number of IL-10+ CD4+ T cells in the mLN (B) after AvCystatin treatment and RSV challenge. Representative data of 2 experiments, 5 mice per group. Error bars indicate SEM. P values reflect Mann-Whitney t-test: * p<0.05, **p<0.01.Total number of FoxP3(TIF)Click here for additional data file.S4 FigP values reflect Mann-Whitney t-test: * p<0.05.Relative expression of MUC5a in mice lungs after the vvG RSV model (A) or primary RSV model (B). Error bars indicate SEM. (TIF)Click here for additional data file.


Ocimum sanctum) were explored and investigated for their diversity and antiphytopathogenic activity against widespread plant pathogens Botrytis cinerea, Sclerotinia sclerotiorum, Rhizoctonia solani and Fusarium oxysporum. 90 fungal isolates, representing 17 genera were recovered from 313 disease-free and surface sterilised plant segments (leaf and stem tissues) from three different geographic locations  during distinct sampling times in consequent years 2010 and 2011 in India. Fungal endophytes were subjected to molecular identification based on rDNA ITS sequence analysis. Plant pathogens such as F. verticillioides, B. maydis, C. coarctatum, R. bataticola, Hypoxylon sp., Diaporthe phaseolorum, Alternaria tenuissima and A. alternata have occurred as endophyte only during second sampling (second sampling in 2011) in the present study. Bi-plot generated by principal component analysis suggested tissue specificity of certain fungal endophytes. Dendrogram revealed species abundance as a function of mean temperature of the location at the time of sampling. Shannon diversity in the first collection is highest in Hyderabad leaf tissues (H' = 1.907) whereas in second collection it was highest from leaf tissues of Delhi (H' = 1.846). Mukteshwar  reported least isolation rate in second collection. Nearly 23% of the total fungal isolates were considered as potent biocontrol agent. Hexane extract of M. phaseolina recovered from Hyderabad in first collection demonstrated highest activity against S. sclerotiorum with IC50 value of 0.38 mg/ml. Additionally, its components 2H-pyran-2-one, 5,6-dihydro-6-pentyl and palmitic acid, methyl ester as reported by GC-MS Chromatogram upon evaluation for their antiphytopathogenic activity exhibited IC50 value of 1.002 and 0.662 against respectively S. sclerotiorum indicating their significant role in antiphytopathogenic activity of hexane extract. The production of 2H-pyran-2-one, 5,6-dihydro-6-pentyl from M. phaseolina, an endophytic fungus is being reported for the first time.Endophytic mycopopulation isolated from India’s Queen of herbs Tulsi ( Endophytic fungi inhabit a unique biological niche and are categorized as highly diverse, polyphyletic group of primarily ascomycetous fungi, capable of colonizing tissues of plants asymptomatically without initiating any disease or overt negative symptoms , 2], , , 5]. 42. 425]. 4Ocimum sanctum is ubiquitous in Indian culture and tradition, called Tulsi in Hindi, . F. FO. sanThe GC-MS analysis of hexane extracts was carried on GC-MS (Agilent Technologies 7890A). For this 1mg of extract was dissolved 1 ml LR-grade dichloromethane . DB-WAX column (30 m × 250μm × 0.25 μm) was used with helium as a carrier gas at a flow rate of 1 ml/min at pressure of 11.654 psi. The GC oven temperature was kept at 120°C for 2 min and programmed to 5°C/min to 130°C for 1 min; 5°C/min to 150°C for 2 min; 2°C/min to 180°C for 3 min; 2°C/min to 200°C for 3 min; 5°C/min to 240°C for 20 min. Splitless injections were done with liquid injection method in this study. A library search was carried out using NIST library.2 placed in such a way that seven are put in periphery and one in the centre of the plate. The control experiment consisted of only methanol solvent in PDA medium , , , 39]..39].The similarity and of endophytic fungal assemblages among both tissues was compared using the following similarity indices:(QS), QS = 2a/(2a + b + c) where ‘a’ is the number of common species in both communities, while ‘b’ and ‘c’ are the number of species specified to each community under investigation, respectively .JS was calculated using the formula: JS = a/(a + b + c) where ‘a’ is the number of common species in both communities, while ‘b’ and ‘c’ are the number of species specified to each community, respectively .O. sanctum different plants parts in distinct sampling time, whilst cluster analysis deduced concurrence between species richness and temperature of geographical locations at the time of sampling. For this, software Unsrcambler X: version 10, CAMO, USA was used . M. Min vitPenicillium sp.), OSHL-5.2 (Alternaria sp.), OSHL-2.1 (M. phaseolina), OSHS-3.1(M. phaseolina), OSML-5.4 (Colletotrichum sp.), OSMS-2.2 (Aspergillus niger), OSMS-2.3 (Meyerozyma guilliermondii), OSDSL-9.8 (Fusarium proliferatum), OSDSS-2.5 (Chaetomium coarctatum), OSDSL-5.6 , OSHSS-5.2 (Sympodiomyces sp.), OSHSS-1.3 (Fusarium proliferatum) OSHSS-2.3 (Diaporthe phaseolorum) endophytic fungal isolates were multiplied on rice medium, and their crude ethylacetate extracts were subjected to intoxicated/poisoned food bioassay to investigate antiphytopathogenic activity of crude fungal extracts against all four phytopathogens. IC50 values of ethylacetate crude extracts of selected thirteen endophytic fungal isolates at five different concentrations was calculated by regression analysis are summarised as Chaetomium coarctatum recorded the highest activity against all phytopathogens with IC50 value of < 1mg/ml ranging from 0.262mg/ml– 0.553 mg/ml. Second best activity was reported with Fusarium proliferatum obtained from Hyderabad with IC50 value ranging from 0.299 to 1.04 mg/ml.Based on the results of dual culture bioassay , followiFusarium proliferatum obtained from Hyderabad exhibited IC50 value of 0.51mg/ml against F.oxysporum which was stronger than another isolate of F. proliferatum recovered from Delhi in second sampling , OSHS-3.1 (M.phaseolina), OSDSS-2.5 (Chaetomium coarctatum), OSHL-5.2 (Alternaria sp.), OSML-5.4 (Colletotrichum sp.) and OSHSS-1.3 (F. proliferatum) were found positive for terpenoids in preliminary phytochemical screening. GC-MS chromatography of hexane extracts of these selected endophytic fungal isolates revealed presence of volatile compounds, fatty acids, and aliphatic constituents. GC-MS chromatogram of M. phaseolina (OSHL-2.1) showed presence of 2H-Pyran-2-one, 5, 6-dihydro-6-pentyl at RT 30.156, hexadecanoic acid at RT 53.017, linoleic acid at RT 64.986 (M. phaseolina (OSHS-3.1) showed presence 9,12-octadecadieonic acid, methyl ester at RT 61.108, trifluoroacetoxy hexadecane at RT 55.970 as major peaks reflecting strain variability (Chaetomium coarctatum (OSDSS-2.5) 2-fluorobenzoic acid at RT 38.644, 2,5-difluorobenzoic acid at RT 38.444, linoleic acid, ethyl ester at RT 64.900 were observed as major peaks (Alternaria sp. (OSHL-5.2) hexane extract, hexadecanoic acid at RT 52.990, 9,15- octadecadienoic acid, methyl ester at RT 61.036 were recorded as major peaks (Fusarium proliferatum (OSHSS-1.3)-major peaks were hexadecanoic acid at RT 52.992, linoleic acid at RT 64.902, oleic acid at RT 65.954 (Colletotrichum sp. (OSML-5.4) major peaks recorded were 7-hydroxy3- coumarin at RT 61.372, 9, 12-octadecadieonic acid at RT 61.052 . Rhizopus oryzae has been reportedly primary causal agent of tuber rot disease in sweetpotato in India and in this study has been isolated from Delhi and Hyderabad in first sampling season . I. IO. sanic fungi , 51], , , , and 7272.Additionally, cluster analysis inferred distribution and abundance of endophytic fungal species from different geographical locations as a function of temperature. It grouped together species richness of Delhi (second sampling) and Hyderabad (first sampling) when temperature ranged from 23–24°C into cluster 1, cluster 2 comprises of Delhi (first sampling) and Hyderabad (second sampling) when temperature ranged from 30–31°C and cluster 3 belongs to endophytic mycobiota of Mukteshwar where temperature range was 14–17°C during both sampling times. This revelation is in accordance with previous studies elucidating higher influence of temperature, precipitation and other climatic factors than geography on endophytic communities of plants , 74]..74].S. Sclerotiorum causes heavy yield losses up to the tune of 40% in Brassicas in India , ,  all the in India . Fusariuin India . Botryti disease . R. solahe world . Differeironment .M. phaseolina (OSHL-2.1) exhibited effective inhibitory activity against B. cinerea, F. oxysporum and R. solani with 4.504 mg/ml, 1.97mg/ml and 1.41mg/ml IC50 values respectively. Whereas hexane extract of M. phaseolina (OSHL-2.1) demonstrated highest activity against S. sclerotiorum with IC50 value of 0.38 mg/ml followed by F. oxysporum with IC50 value of 0.867mg/ml. Preliminary identification of bioactive metabolites were conducted on crude hexane extracts.GC-MS chromatogram of hexane extract of M. phaseolina illustrated presence of 2H-pyran-2-one, 5, 6-dihydro-6-pentyl at RT 30.152 and palmitic acid, methyl ester at RT 53.017, while that of C. coarctatum (OSDSL-2.5) illustrated Oleic acid at RT 61.675. Present study for the time reports antifungal activity of extracts of M. phaseolina and C. coarctatum. Antifungal activities of above mentioned pure compounds against plant pathogens resulted in remarkable revelation. Oleic acid methyl ester, palmitic acid methyl ester and 5, 6 Dihydro-2H-pyran-2-one-6 pentyl exhibited strong inhibitory activity against S. sclerotiorum with 0.547mg/ml, 0.662mg/ml and 1.002 mg/ml IC50 value while against F. oxysporum it was 0.27 mg/ml, 0.264 mg/ml and 0.27mg/ml respectively. Thereby, buttressing the fact that 2H-pyran-2-one, 5,6-dihydro-6-pentyl and palmitic acid, methyl ester are bioactive metabolites synthesised by endophytic fungus M. phaseolina which are responsible for its antiphytopathogenic activity against S. sclerotiorum. Antifungal activity of 6-Pentyl-2H-pyran-2-one and its analogs has been well documented in literature . R. RM. phalium spp . Anotheric fungi . The pro studies . Hexadecalbicans . The majNCBT 196 . Another. solani .Bipolaris maydis, Rhizoctonia bataticola and Chaetomium coarctatum in endophytic state harbored inside O. sanctum for the first time. In present study metabolites produced by M. phaseolina recovered from Hyderabad in first collection has exhibited promising antifungal activity against S. sclerotiorum and F. oxysporum broad spectrum phytopathogens. Metabolites originated from fungal endophytes hold promise to be further developed as greener and safer biocontrol agent in crop disease management. It can be concluded that fungal endophytes harbored inside leaf and stem tissues of Ocimum sanctum collected from different geographical locations in different sampling times hold great promise not only as biocontrol agents against broad spectrum and economically significant phytopathogens, but also as sustainable resource of novel antifungal secondary metabolites.The study reports noted plant pathogens such as S1 Fig(TIFF)Click here for additional data file.S1 Table(DOCX)Click here for additional data file.S2 Table(DOCX)Click here for additional data file.


Cancer patients are at risk for severe complications related to the underlying malignancy or its treatment, and therefore usually require admission to ICUs.To evaluate the clinical characteristics and outcomes in this subgroup of patients.Analysis of two prospective cohorts of cancer patients admitted to ICUs. We used multivariable logistic regression to identify variables associated with hospital mortality.P <0.001 for all analyses). ICU (47 % vs. 27 %) and hospital (63 % vs. 38 %) mortality rates were also higher in patients with complications (P <0.001). Chemo/radiation therapy-induced toxicity (6 %), venous thromboembolism (5 %), respiratory failure (4 %), gastrointestinal involvement (3 %) and vena cava syndrome (VCS) (2 %) were the most frequent complications. In multivariable analysis, the presence of cancer-related complications per se was not associated with mortality (odds ratio (OR) = 1.25 ). However, among the individual complications, VCS (OR = 3.79 (1.11-12.92)), gastrointestinal involvement (OR = 3.05 (1.57-5.91)) and respiratory failure (OR = 1.96 (1.04-3.71)) were independently associated with worse outcomes.Out of 2028 patients, 456 (23 %) had cancer-related complications. Compared with those without complications, they more frequently had worse performance status (PS) (57 % vs. 36 % with PS ≥2), active malignancy (43 % vs. 5 %), need for vasopressors (45 % vs. 34 %), mechanical ventilation (70 % vs. 51 %) and dialysis (12 % vs. 8 %) (The prognostic impact of cancer-related complications was variable. Although some complications were associated with worse outcomes, the presence of a severe acute cancer-related complication per se should not guide decisions to admit a patient to the ICU.


Apelin (APLN) is a powerful inotrope, is widely expressed by the cardiovascular system with its receptor APJ-R, and should be considered as an alternative noncatecholaminergic support.Dobutamine (DOB) is the actual recommended b-adrenergic inotropic drug to support sepsis-induced myocardial dysfunction when cardiac output index is still low after preload correction. In this context, DOB cardiovascular response predicts outcome in septic shock. Alternative supportive and safer therapies are however mandatory because:Perform a comparative evaluation of APLN-13 (APLN active peptide) vs DOB in terms of hemodynamic efficacy, cardioprotection and outcome in a model of “sepsis-induced cardiac dysfunction”.E. Coli 055:B5, 10-12mg/kg intraperitoneal).A rat model of LPS-induced myocardial dysfunction .0-6-18h.in vivo: echocardiography, final hemodynamics; urine output, weight and plasma volume variation, survival study, ex vivo: Langendorff dP/dt, in vitro: APJ-R and beta1-adrenergic receptor (AR) myocardial expressions, Pi3K/Akt/GSK3/mTOR activation-expression profiles, cTnI  and cleaved caspase-3 (apoptosis).Both drugs restored LPS-induced fall of left ventricular ejection fraction (LVEF) with dominant chronotropic impact and mean arterial pressure (MAP) restoration for DOB, and lower peripheral vascular resistances (PVR) for APLN. APJ-R but not beta1 AR myocardial expressions were upregulated by LPS challenge (p< 0.05). The deepness the induced LVEF drop, the higher the dP/dt response to APLN but not to DOB (p< 0.05). In 18h LPS-challenged hearts, Langendorff assays peak dP/dt responses were 3nM and 100nM for APLN and DOB, respectively (p< 0.05). Combining fluid resuscitation with APLN infusion declined urine output (UO) with plasma volume (PV) expansion, whereas DOB induced less PV expansion but more UO (p< 0.05). Survival proportions were clearly distinctive (p< 0.05).APLN further dampened down LPS-induced overphosphorylation/inhibition of Pi3K/Akt/mTOR/GSK3 and reduced injury/apoptosis (i.e. cTNI and cleaved caspase-3 expressions).APLN potentially offers distinctive mechanisms of hemodynamics, cardioprotective effects, and survival benefits, over DOB. Chemical optimization of APLN-13 with more extensive preclinical data, would pave the way for first phase clinical trials.


Due to increasing resistance, alternatives to mupirocin and chlorhexidine for decolonization of MRSA carriage need to be evaluated.®) vs placebo in eliminating MRSA carriage at day 28 (D28) after the end of treatment.To evaluate the efficacy of polyhexanide  or placebo  applied to the anterior nares and skin for 10 days. The primary outcome was MRSA decolonization at D28 assessed by both intention-to-treat  and per-protocol (PP) analysis . Secondary outcomes included MRSA decolonization according to nasal MRSA carriage, safety and emergence of resistance.vs 22/75 (29.3%) in group P were MRSA-free at D28 . PP analysis confirmed the results with 19/53 (35.8%) decolonized polyhexanide-treated patients vs 17/56 (30.4%) in the placebo arm . In the subgroup of MRSA nasal carriers, PP analysis showed that 6/15 (40.0%) patients in group I vs 2/11 (18.2%) in group P were decolonized (P=0.40). Nine serious adverse events occurred in group I vs 12 in group P; none was attributable to study medication. Emergence of polyhexanide resistance was not observed.Of 2590 patients screened, 146 patients  were randomized between January 2011 and July 2014. Primary outcome was missing for 11 (7.5%) patients. ITT analysis showed that 24/71 (33.8%) patients in group I This study suggests that under real-life conditions a single polyhexanide decolonization course is marginally effective in eradicating MRSA carriage.C. Landelle: None declared, E. Von Dach: None declared, T. Haustein: None declared, A. Agostinho: None declared, G. Renzi: None declared, A. Renzoni: None declared, D. Pittet: None declared, J. Schrenzel: None declared, P. François: None declared, S. Harbarth Grant/Research support from: a peer-reviewed research grant funded by Pfizer, Consultant for: the advisory boards of Destiny Pharma, bioMerieux, Novartis, and DaVolterra


COBI, a PK enhancer with no ARV activity is a more selective cytochrome P450 (CYP)3A inhibitor than ritonavir (RTV), does not induce CYP isozymes, and thus has less potential for drug-drug interactions. COBI boosts DRV PK as effectively as RTV in healthy volunteers.This 48-week, phase IIIb, open-label, single-arm, US multicentre study (NCT01440569) included HIV-infected treatment-nave and experienced adults with no DRV RAMs, viral load (VL) ≥1000 c/mL, eGFR ≥80 mL/min and genotypic sensitivity to investigator-selected N[t]RTIs. Patients received DRV/COBI 800/150 mg qd (as single agents) plus two fully active N[t]RTIs. The primary endpoint was any treatment-emergent grade 3 or 4 AEs through Week 24. We report 48-week safety, efficacy and PK/PD results in treatment-nave patients.10 c/mL and CD4+ 370 cells/mm3. Treatment-emergent grade 3 or 4 AEs regardless of causality were reported in 21 (7%) patients. AEs regardless of causality  were: diarrhoea (27%), nausea (23%), URTI (15%) and headache (12%). Sixteen (5%) patients had AEs leading to study drug discontinuation, most frequently rash (three patients), hypersensitivity and nausea (two patients each). Consistent with the known inhibition of tubular creatinine secretion by COBI, there was a mean increase from BL in serum creatinine by week 2 (0.09 mg/dL), remaining stable through week 48 (mean 0.10 mg/dL increase from BL). At week 48, 83% of patients achieved VL<50 c/mL; FDA Snapshot); median increase in CD4+ was 169 cells/mm3. Eight patients met the criteria for resistance testing. M184V was detected in one pt receiving FTC. New primary RAMs were not detected in the other seven patients. The mean population PK-derived DRV AUC24h was 100,620 ng.h/mL and C0h 2,105 ng/mL (n=281). There were no clinically relevant relationships between DRV exposure and virologic response, AEs or laboratory parameters.Of 313 ITT patients, 295 were treatment-nave (94%). In the treatment-nave cohort, 90% were male, 60% white and 294 (99.7%) received a TDF-containing regimen. Median baseline (BL) VL was 4.8 logThe DRV PK of DRV/COBI was consistent with historical data for DRV/RTV. DRV/COBI 800/150 mg qd plus two N(t)RTIs had an 83% response and was well tolerated through Week 48. These results are similar to published data for DRV/RTV 800/100 mg qd, and support the use of DRV/COBI 800/150 mg qd in treatment-nave patients.


Methylobacter luteus, Methylobacter whittenburyi, Methylosarcina fibrata, Methylomicrobium agile, and Methylovulum miyakonense were generated. The strains represent aerobic methanotrophs typically isolated from various terrestrial ecosystems.Genome sequences of  Methylobacter whittenburyi (formerly “Methylobacter capsulatus” = UCM-B-3033), and Methylomicrobium agile (ATCC 35068) are methanotrophic bacteria commonly found in sediment samples from wetlands  have typically been obtained from meadows, dry hay, and cow mouth samples (–Methylovulum miyakonense HT12T (= ATCC BAA-2070) was isolated from a forest soil (Methylosarcina fibrata AML-C10T (= ATCC 700909) was isolated from a landfill site , using the Illumina  and/or P17–Prodigal  and GeneProdigal . AdditioProdigal  and MaGeProdigal  platformM. miyakonense HT12T genome (pxmABC)  or urea (urtABCDE/ureABCDEFG) as the sole source of nitrogen. M. miyakonense HT12T, M. luteus 98, and M. whittenburyi UCM-B-3033 possess the key genetic elements for nitrogen fixation (nifKDHWENX).Genome statistics and predicted core metabolic pathways are shown in T genome . A funct(pxmABC)  was founrogenase . Two typ protein . M. luteeductase  were ideMethylobacter spp.) produce cysts (M. luteus 98 (Many methanotrophic species (including ce cysts . We wereuteus 98 , 30. Twouteus 98  were ideThe genome sequences have been deposited in GenBank under the accession numbers listed in