Patients with locally advanced and high-risk prostate cancer (LAPC) are prone to experience biochemical recurrence despite radical prostatectomy (RP). We evaluated feasibility, safety and activity of a neoadjuvant chemohormonal therapy (NCHT) with 3-weekly full dose docetaxel and complete androgen blockade (CAB) in locally advanced and high-risk prostate cancer patients (LAPC) undergoing RP.2) followed by RP. Primary endpoints were biochemical (PSA) and local downstaging. Secondary endpoints included toxicity and operability assessments, pathological complete response (pCR), time to PSA progression, 5-year biochemical recurrence free survival (bRFS) and overall survival (OS).Patients (n = 30) were selected by Kattans’ preoperative score and received trimestral buserelin 9,45 mg, bicalutamide 50 mg/day and 3 cycles docetaxel (75 mg/mMedian baseline PSA was 25.8 ng/ml (2.1–293), and the predicted probability of 5-year bRFS was 10% (0–55). NCHT induced PSA-reduction was 97.3%  and post-RP 96.7% of patients were therapy responders, with undetectable PSA-values. Post- vs. pretreatment MRI indicated a median tumor volume reduction of 46.4% . A pathological downstaging was observed in 48.3%. Severe hematologic toxicities (≥CTC3) were frequent with 53.8% leucopenia, 90% neutropenia and 13.3% febrile neutropenia. RP was performed in all patients. While resectability was hindered in 26.7%, continence was achieved in 96.7%. Pathologic analyses revealed no pCR. Lymph node- and extracapsular involvement was observed in 36.7% and 56.7% with 33.3% positive surgical margins. After a median of 48.6 (19.9-87.8) months, 55.2% of therapy responders experienced PSA-recurrence. The estimated median time to PSA-progression was 38.6 months (95%CI 30.9-46.4) and 85.3 months (95%CI 39.3–131.3) for OS. The 5-year bRFS was improved to 40%, but limiting for interpretation adjuvant treatment was individualized.NCHT is feasible despite high hematotoxicity, with excellent functional results. Significant downstaging was observed without pCR. NCHT seems to improve the cohort adjusted 5-year bRFS, but clinical value needs further investigation in randomized trials. Patients with locally advanced and high-risk prostate cancer (LAPC) are prone to experience biochemical recurrence despite curative-intended radical prostatectomy (RP). Predictor variables are PSA-value, clinical stage and Gleason score ,2. In orIn castration-resistant prostate cancer patients (CRPC), docetaxel chemotherapy (D) is standard of care due to prolonged survival in randomized phase III trials -11. SubsSince androgen-dependent and -independent cell subpopulations may coexist, neoadjuvant strategies combining docetaxel and complete androgen blockade (CAB) were investigated, still resulting in deficient pCR rates of 3-6% ,16. In f2), over a period of 3 cycles (9 weeks), may exhibit superior activity to downstage tumors and improve oncological outcome. Therefore, we prospectively determined feasibility, safety and activity of this presurgical short-term combination therapy in 30 LAPC patients undergoing RP.The combination of CAB with a 3-weekly full dose regimen of docetaxel  patients completed all 3 cycles. Three patients (10%) discontinued NCHT; one withdrew consent after one cycle, and two interrupted due to NCHT-unrelated severe AE ) after two cycles of docetaxel. Bicalutamide was continued up to RP except for the patient with PAOD. Dose reductions of docetaxel were necessary in two patients (6.7%) and a treatment deferral in one patient (3.3%). Adverse events occurring in ≥10% of patients are presented in Table All patients n = 30) underwent RP with histopathological analyses after a median interval of 70 days from day 1 (range 61–143). Median removed lymph nodes were 16 (range 5–39). Surgical variables are depicted in Table 0 underweLong time functional outcome, following a median of 48.6 months (range 14.1-87.8), revealed continence in 96.7%. Before treatment 86.7% (n = 26) patients were potent. After RP 15.4% had spontaneous erections (E3-E4), 11.5% were potent with erection aids and 19.2% had tumescence sufficient for sexual activity but not for vaginal penetration (E1-E2).All patients (n = 30) demonstrated a PSA reduction following NCHT with a median decline of 97.3%  , without upstaging. Prostate volume was reduced in all cases with a significant median decrease of 37.1% . Similarly a significant median tumor volume reduction of 46.4%  was observed in 95.7% (n = 22) of patients harboring measurable tumors, while one progressed (4.3%). Pathological downstaging, comparing initial T-stages on MRI (n = 29) with histopathological stages revealed downstaging in 48.3%  and upstaging in 13.8% . Histopathological analyses revealed no pathological complete response (pCR), and only one (3.3%) pathological minimal residual disease (pMRD) with a tumor volume of <5%.After a median FU of 48.6 months (range 19.9-87.8), of the 29 treatment responders 55.2% experienced a biochemical progression. The estimated median time to PSA progression was 38.6 months (95% CI 30.9-46.4) with a 5-year biochemical recurrence free survival of 40%  patients over a mean of 19 months (range 11–28) due to pathological R1N1 (n = 2) or R0N1 (n = 5) stages. Adjuvant radiation therapy (RT) was performed in 3 (10%) patients due to positive surgical margins without lymph node involvement (R1N0). No adjuvant RT was performed in 3 patients despite a R1N0 stage in favor a deferred RT in case of biochemical recurrence and in 4 patients with a R1N1 stage.At the time of this analysis 26 (86.7%) patients were still alive. Two patients (6.7%) experienced a PC related death. The estimated median OS in the complete cohort was 85.3 months (95%CI 39.3–131.3) , combined with CAB in LAPC patients undergoing RP. Although presurgical docetaxel-based regimens were investigated earlier, FU data are mostly short-term Table . In addiWith respect to non-hematologic toxicities, we observed the known profile of AE during palliative or neoadjuvant D ± ADT. The incidence of severe AE was low (7%) when compared to weekly D monotherapy (21-53%) or weekly D + CAB (21-26%) ,13,15,16Surgical variables are in accordance with literature. Resectability was partly hindered due to callosity as reported earlier for neoadjuvant docetaxel or CAB ,13,15-19Several trials used a pCR rate of 10-40% as primary endpoint, but failed ,13,15-17With respect to downstaging we observed a partial PSA response in 100% and a complete PSA response in 13%. The PSA-value reduction was 97%, similar to weekly D + CAB or D + EMP, but superior to docetaxel monotherapy with 64% PSA-decline and 24% partial response ,16,21. INeoadjuvant trials with long-term FU and relevant recurrence rates are rare. Using D + ADT + EMP a recurrence rate of 58% was observed after a FU of 53 months . SimilarAlthough our trial gives additional clinical information and confirms significant clinical activity of neoadjuvant D + CAB, our study has certain limitations especially with respect to bRFS. Adjuvant treatment was not uniformly defined. This is in accordance with literature, but contrasts with other trials that excluded adjuvant therapy until progression ,15,16,23To our knowledge we are the first to present a long time FU of a phase II trial, investigating a short-term neoadjuvant therapy with 3-weekly full dose docetaxel, combined with CAB in locally advanced and high-risk PC patients scheduled for RP. The regimen is feasible, despite a high rate of severe neutropenia and leucopenia. Significant clinical activity was observed, despite absence of pCR. Surgical variables revealed an excellent functional outcome. Finally a precariously 30% improvement of the 5-year bRFS was achieved.This single-center non-randomised phase II trial was conducted at the Department of Urology, Klinikum rechts der Isar, Technische Universität München. The study was approved by the institutional review board and the regulatory authorities and was performed in accordance with the ethical standards of the Declaration of Helsinki. All subjects gave written informed consent. Main eligibility criteria were adenocarcinoma of the prostate, absence of metastatic disease and a biochemical recurrence risk of >40% within 5 years, according to Kattan’s preoperative nomogram .Primary objectives were downstaging effects, reflected by PSA-value decrement and local tumor reduction by clinical and pathological criteria. Secondary objectives were the pCR and pathological minimal residual disease (pMRD) rate, the toxicity profile and operability, along with time to PSA progression, 5-year biochemical recurrence free survival (bRFS) and overall survival (OS).2) on days 8, 29 and 50 in combination with prednisolone 10 mg/day p.o. starting on day 9. Dexamethasone (8 mg p.o.) was administered 12 hours before and after docetaxel as well as 60 min before. A dose reduction of docetaxel to 60 mg/m2 and a treatment deferral of 21 days were allowed. All patients underwent open retropubic RP including extended bilateral pelvic lymph node dissection up to the aorta. Adjuvant radiation and hormonal therapies were individualized according to pathological results . NCHT was aborted in case of unacceptable toxicities, progression or on patients’ request.Presurgical treatment consisted of CAB comprising bicalutamide 50 mg/day p.o. starting day 1 until the day before RP with trimestral buserelin 9.45 mg s.c. on day 3 and three cycles of docetaxel chemotherapy  or magnetic resonance imaging (MRI) of the pelvis. Pelvic lymph nodes were considered non-metastatic if the maximum diameter did not exceed 1.5 cm. Local clinical downstaging was assessed by MRI of the prostate with endorectal coil  before and after NCHT. T2-weighted sequences in three planes, axial T1-weighted and a dynamic-contrast-enhanced sequence were analyzed independently by a board certified radiologist. Median prostate and tumor volumes were calculated ,25-27. TSurgical variables were recorded implying the functional outcome. Continence was defined as the use of no pads and erectile function using the erection hardness score (E0-E4) ,29. AdveThis trial was conducted as an intent to treat approach . Sample The authors have no actual or potential conflict of interest in relation to this article to declare. The study was supported by Sanofi-Aventis, Frankfurt, Germany.MT and HRK analyzed data and wrote the manuscript, UT contributed to concept design, JG and MR directed the study and contributed to data interpretation, MT, TH, MMH, TM and MA performed the research and contributed to data collection, BJK and ME evaluated imaging datasets, KH edited the manuscript. All authors read and approved the final manuscript.


SOD1) in rodents has provided useful models for studying the pathogenesis of amyotrophic lateral sclerosis (ALS). Microglia have been shown to contribute to ALS disease progression in these models, although the mechanism of this contribution remains to be elucidated. Here, we present the first evidence of the effects of overexpression of mutant (TG G93A) and wild type (TG WT) human SOD1 transgenes on a set of functional properties of microglia relevant to ALS progression, including expression of integrin β-1, spreading and migration, phagocytosis of apoptotic neuronal cell debris, and intracellular calcium changes in response to an inflammatory stimulus.Overexpression of mutant copper/zinc superoxide dismutase (TG SOD1 G93A but not TG SOD1 WT microglia had lower expression levels of the cell adhesion molecule subunit integrin β-1 than their NTG control cells , resulting in decreased spreading ability, with no effect on ability to migrate. Both TG G93A and TG WT microglia had reduced capacity to phagocytose apoptotic neuronal cell debris , and 26.9 ± 2.8% for NTG (WT) cells). Extracellular stimulation of microglia with ATP resulted in smaller increase in intracellular free calcium in TG G93A and TG WT microglia relative to NTG controls , and 0.37 ± 0.05 μM for NTG (WT) microglia).SOD1 transgenic mice have a reduced capacity to elicit physiological responses following tissue disturbances and that higher levels of stimulatory signals, and/or prolonged stimulation may be necessary to initiate these responses. Overall, resting mutant SOD1-overexpressing microglia may have reduced capacity to function as sensors of disturbed tissue/cellular homeostasis in the CNS and thus have reduced neuroprotective function.These findings indicate that, under resting conditions, microglia from mutant  SOD1) i) was measured before, during, and after stimulation with 10 μM ATP. Stimulation with ATP increased [Ca2+]i and withdrawal of ATP resulted in a steady reduction of the [Ca2+]i to almost baseline levels in microglia of all genotypes , 0.14 ± 0.01 μM for TG G93A, 0.12 ± 0.01 μM for NTG (WT) and 0.13 ± 0.01 μM for TG WT. The first 10 s of ATP stimulation resulted in an initial rapid increase of [Ca2+]i in NTG microglia, followed by a steady reduction of [Ca2+]i during the remaining 50 s of ATP challenge. However, TG microglia displayed a different pattern of [Ca2+]i changes; the initial peak in [Ca2+]i was absent in both TG G93A and TG WT microglia  calculations quantified this pattern of [Ca2+]i changes, and showed areas of 19.1 ± 1.8 μM*s for NTG (G93A) versus 14.4 ± 2.2 μM*s for TG G93A microglia, p = 0.0176, Student's t test, and 14.6 ± 1.5 μM*s for NTG (WT) versus 11.9 ± 1.6 μM*s for TG WT cells, not significantly different  cells, p = 0.003, Student's t test, and 0.24 ± 0.03 μM for TG WT versus 0.37 ± 0.05 μM for NTG (WT) microglia, p = 0.0123, Student's t test  is involved in diverse cellular functions including cellular adhesion to extracellular matrix, focal adhesion, migration, actin polymerisation/assembly, and regulation of the cell cycle -50. We oin vivo after injection of toxin ricin into rat facial nerve  at 60 000 cells/cm2 on poly-L-lysine (Sigma) coated coverslips. For the purification of microglia, the confluent cultures were subjected to shaking and mild trypsinisation i in NTG (G93A) and TG G93A microglia following extracellular stimulation with 1 mM ATP for 1 min. The period of ATP challenge is indicated with the bar. (C) Area-under-the-curve (AUC) values (μM*s) for NTG (WT), TG WT, NTG (G93A) and TG G93A microglia. Data are mean ± SEM; Student's t test, *p = 0.0201 versus NTG (WT) microglia. (D) [Ca2+]i values (μM) recorded for NTG (WT), TG WT, NTG (G93A) and TG G93A microglia at 130 s of recording . Data are mean ± SEM; Student's t test, *p = 0.0381 versus NTG (WT) and **p = 0.0015 versus NTG (G93A) cells, n = three NTG (WT) mice (14 cells) and three TG WT mice (18 cells) from two litters, three NTG (G93A) mice (43 cells) and three TG G93A mice (38 cells) from two litters.Click here for file


Patients with advanced pancreatic adenocarcinoma have a poor prognosis, and to date, no treatment method has had a significant impact on the disease. In general, the mean overall survival time of such patients receiving conventional chemotherapy and radiotherapy is <6 months. In the present case report, a patient with advanced pancreatic adenocarcinoma experienced a longer progression-free survival (PFS) of >19 months, following cytokine-induced killer (CIK) cell therapy. To the best of our knowledge, no study has previously described such a beneficial effect on patients only receiving CIK cell immunotherapy. Based on these findings, CIK cell therapy may be a potential treatment regimen that is capable of leading to an improved prognosis in certain patients with advanced pancreatic adenocarcinoma. The medium was replaced by fresh IL-2- and IFN-γ-containing medium every 5 days. At day 10, CIK cells were harvested and their phenotypes were analyzed. All products were free of bacterial, mycoplasma and fungal contamination. The endotoxin level was <5 EU. Phenotypic analysis of autologous CIK cells in the patient prior to culture and following 10 days of cultivation demonstrated that the percentages of CD3+, CD3+CD4+, CD3+CD8+, CD3+CD56+ and CD25+ cell subsets increased from 45.40±5.21, 29.08±4.86, 18.80±5.45, 3.77±1.665 and 12.51±4.01% to 90.06±9.22, 44.50±8.18, 38.40±4.19, 15.21±4.53 and 31.75±5.87%, respectively (P<0.01). However, the percentages of CD3+/16+/56+, CD14+ and CD20+ cell subsets decreased from 14.73±3.54, 16.46±6.06 and 11.19±3.18% to 6.78±1.91, 5.87±2.09 and 7.84±2.53%, respectively (P<0.05). The total number of CIK cells in one cycle is ∼ 5×109. From November 10, 2010 to June 27, 2011, the patient recieved 4 cycles of CIK cell immunotherapy and 2 million units of IL-2 from day 1–5 per infusion of CIK cells. In the period of CIK cell therapy, no adverse reactions were observed. Following 4 cycles of CIK cell immunotherapy, the abdominal contrast-enhanced CT reexamination demonstrated that the low-density nodule reduced significantly  reexamination after one month revealed that a low-density nodule had emerged at the resection margin . Cancer-ficantly . The canficantly  comparedficantly . From MaIn the field of oncology, pancreatic cancer remains a fatal disease with a poor prognosis, thus presenting a challenge for oncologists ,13. In pin vitro, with stronger antitumor activity in a broad spectrum of tumors  was prolonged by 27 months compared with the control arm  and significantly improved the mOS (Presently, immunotherapy has become the fourth treatment modality for malignant tumors, and may be a promising approach for pancreatic cancer –10. Certf tumors ,17. In af tumors ,19. The f tumors . In part the mOS . HoweverIn conclusion, we describe the case of a 77-year old female with pancreatic cancer who underwent surgery, although the bilateral margins were positive. After one month, imaging examination revealed that a low-density nodule had emerged at the resection margin. Considering the patient’s poor condition, low KPS scores and inability to tolerate the side-effects of chemotherapy and/or radiotherapy, the patient was treated with CIK cell immunotherapy. The patient achieved a PFS of >19 months, which was longer than that demonstrated by using chemotherapy and/or radiotherapy. In conclusion, CIK immunotherapy may be an effective treatment method for advanced pancreatic cancer, particularly for elderly patients.


Subjects were 13 elite male kayakers, aged 20.0 ± 3 years, with average body mass of 75.0 ± 7.9 kg and 177.3 ± 7.1 cm height and with a VO2max of 58.3 ± 7.8 mL·kg−1·min−1. The nonathletes were 7 men, aged 18.2 ± 1.1 years, body mass of 81.3 ± 13.8 kg, and 171.9 ± 4.5 cm height. Blood samples were collected after six weeks of offtraining and before the start of a new training season. PB leukocyte populations were determined by flow cytometry. Cytokine levels were quantified by ELISA. When nonathletes were compared with the kayakers, the latter exhibited lower plasma concentrations of IL-1β, IL-18, and IFN-γ as well as a lower concentration of IL-1ra. Positive correlations between IL-18 and B cells in the athletes were also found. These results seem to reinforce the anti-inflammatory role of regular training.Regular moderate exercise has been shown to have anti-inflammatory effects that help prevent several chronic diseases. However, the effects of chronic training an elite athletes have not been the focus of much research. This study aimed to determine whether there were differences in cytokine levels (IL-1 Regular exercise has been associated with protection against mortality from all causes, primarily to protect against atherosclerosis, type 2 diabetes, colon and breast cancer . In addiγ) production. It has been suggested that this may be an important mechanism by which exercise leads to a depressed immune function [β, IL-12, IFN-γ, TNF-α, and MIP-1β decreased in highly trained swimmers throughout a training season. It has been proposed that the anti-inflammatory effects of exercise [Prolonged exercise, on the other hand, has been associated with a transient depression in the immune system . Both prfunction , 14. Ourfunction  that theexercise , possiblβ, IL-1ra, IL-6, IL-10, IL-18, IFN-γ, and TNF-α) and leukocyte subpopulations by comparing a group of elite kayakers with age-matched nonathletes.At rest, the immune function of athletes seems to be similar to nonathletes , 11. How2max of 58.3 ± 7.8 mL·kg−1·min−1. The control group consisted of seven healthy men who did not practiced regular physical activity and with a mean age of 18.2 ± 1.1 years, body mass of 81.3 ± 13.8 kg, and 171.9 ± 4.5 cm height. After oral and written explanation, written consent was obtained and the experimental protocol approved by the Ethics and Human Subjects Review Board at the Faculty of Sports Science and Physical Education, University of Coimbra.The elite athlete sample was comprised of 13 elite kayakers, with at least 3 years of kayak training at elite level, mean age 20 ± 3 years, with an average body mass of 75.0 ± 7.9 kg, 177.3 ± 7.1 cm in height, and a VOFor this study, a venous blood sample of the athletes and controls was taken in November, during a specific training period where kayakers worked out with reduced loads and intensities, after an off season transitional period of six weeks, which would be the start of a new training season.For this study, 20 mL of peripheral blood was collected by venipuncture of the antecubital vein, after 48 hours of rest from the last training session. Peripheral blood leukocytes levels, including neutrophils, lymphocytes, and monocytes, were determined using a cell counter .+) and their T-cell subpopulations CD3+CD4+ and CD3+CD8+, B lymphocytes (CD19+), and natural killer cells (NK) (CD3−CD56+). Staining of the cells was performed using the LYMPHOGRAM kit  according to the manufacturer's instructions. The stained cells were analyzed on a FACScan flow cytometer  using CellQuest software (BD Biosciences). Data were analyzed using the software “Infinicyt” .Lymphocyte subsets were determined by flow cytometry, including the total number of T lymphocytes .The concentrations of cytokines IL-1, IL-1RA, IL-6, IL-10, IL-18, TNF-U test to detect differences between athletes and controls and the Spearman correlation test to study the correlations between variables. The value of significance was set at P < 0.05. Statistical analysis was performed using the SPSS software for Mac (version 19.0).For this study we considered the mean and standard deviation values. Because according to the Shapiro-Wilk the samples were not normally distributed, we used the Mann-Whitney β, IL-1ra, IL-18, and IFN-γ plasma concentrations were found for kayakers when compared to the nonathletes. The same was observed for the NK cell population. No differences for IL-10 and IL-6 plasma concentrations were found. The total number and percentage of leukocytes, monocytes, granulocytes, T lymphocytes, and their subpopulations and B lymphocytes did not differ between groups. A compilation of the data is presented in  Lower levels of IL-1ρ = 0.67, P < 0.05) and IL-18 . Negative correlations were found between the % CD3+ and IFN-γ  and the total number of CD3+CD8+ cells and IL-1β . Correlations were also found between the % of B cells and IL-18  and IFN-γ . The total number of B lymphocytes also correlated with IL-18 . All correlation results are shown in Using the Spearman test (Rho), correlations between cells  and plasma interleukin concentrations were found for the kayakers. Leukocytes (WBC) positively correlated with IL-1ra  observed during the training phases of increased volume and intensity [In the present study, NK cells were lower in the kayakers than in the control group. A lower value of CD3ntensity . The effntensity , 21. Manntensity , 23. Thentensity , 21 but + cytotoxic lymphocytes and NK cells were the only cell types producing IFN-γ. Currently there is evidence that other cells such as B cells, NKT cells, and professional antigen-presenting cells (APCs) secrete IFN-γ [γ production is controlled by cytokines secreted by APCs, most notably IL-12 and IL-18. These cytokines serve as a bridge to link infection with IFN-γ production in the innate immune response [γ production. In our study, IL-18 concentration correlated positively with both the percentage and absolute numbers of B cells (CD3−CD19+) (+CD56−) preventing the formation of mature marginal centers [ Initially it was believed that the T helper I (Th1), CD8te IFN-γ . IFN-γ presponse , 25. In 3−CD19+) . This re centers . In anot centers . Our res centers . A signi centers . On the  centers –32.β and CD3+CD8+ cells was also found. The CD3+CD8+ cells are normally exposed during an inflammatory response to IL-1β, IL-6, and IL-23 that arise especially in ischemic lesions [ In the kayakers, a significant inverse correlation between IL-1 lesions . It has  lesions . Recent  lesions . Accordi lesions , the lonβ, IL-18, and IFN-γ that are proinflammatory cytokines. These results appear to demonstrate the anti-inflammatory effects that regular training exercise practice seems to induce and could explain the beneficial effects of long-term regular exercise, manly in preventing chronic low grade inflammation and its consequences. As stated by Gleeson et al. [ When comparing athletes before a training period with a control group of not physically active persons, it was observed that athletes had a lower concentration of IL-1n et al. , in the


The results indicate that the sintering temperature obviously influences the structural and electrical properties of the sample. For the 0.98NKN-0.02BZT ceramics sintered at 1,080°C to 1,120°C, the bulk density increased with increasing sintering temperature and showed a maximum value at a sintering temperature of 1,090°C. The dielectric constant, piezoelectric constant [d33], electromechanical coupling coefficient [kp], and remnant polarization [Pr] increased with increasing sintering temperature, which might be related to the increase in the relative density. However, the samples would be deteriorated when they are sintered above the optimum temperature. High piezoelectric properties of d33 = 217 pC/N, kp = 41%, dielectric constant = 1,951, and ferroelectric properties of Pr = 10.3 μC/cm2 were obtained for the 0.98NKN-0.02BZT ceramics sintered at 1,090°C for 4 h.Lead-free 0.98(Na BZT has the advantage of exhibiting improved piezoelectric properties. However, it has a low Curie temperature (about 100°C), which limits its practical application as a piezoelectric material. In view of the high Curie temperature of NKN ceramics, the NKN-BZT binary system is of much value as a piezoelectric material [0.5K0.5)NbO3-0.02Ba(Zr0.52Ti0.48) [0.98NKN-0.02BZT] solid solution by a conventional ceramics technique, and the influence of sintering temperatures on the dielectric and piezoelectric properties of the 0.98NKN-0.02BZT ceramics was investigated.Lead-based perovskites have been extensively used in industries as sensors, actuators, and transducers due to their outstanding electrical properties. However, the PbO in these materials presents an environmental problem. The studies are now focused on discovering an alternative for lead-based materials. Potassium sodium niobate NbO systems -3. When material . In this0.5K0.5)NbO3-0.02Ba(Zr0.52Ti0.48) ceramics. For specimens prepared by the conventional mixed oxide method from Na2CO3, K2CO3, Nb2O5, BaCO3, ZrO2, and TiO2 as the staring materials, these powders were separately dried in an oven at 100°C for 4 h. They were ball-milled for 24 h using zirconia balls in alcohol. After drying at 110°C for 24 h, the powders were calcined at 850°C for 5 h. The calcined powders were pressed into disk samples of φ = 12 mm. The samples were sintered at 1,080°C to 1,120°C for 4 h. After the samples were polished up to 1.0-mm thick, Ag paste was screen-printed on the surfaces as electrodes and then fired at 400°C for 10 min. We used X-ray diffraction [XRD] and scanning electron microscopy [SEM] to analyze the crystalline and microstructures. The dielectric properties were measured using an LCR meter . Hysteresis loops of the samples were measured by a Sawyer-Tower circuit. The samples were poled under a DC field of 4 kV/mm for 20 min. The d33 was measured by a d33 meter . The kp was calculated by measuring the antiresonance and resonance frequencies. The relative density of the sintered samples was measured by the Archimedes method.The chemical molecular formula used in this experiment for the perovskite ceramics with  complex A-sites and  complex B-sites is 0.98 mode. Figure Pr is 5.8 μC/cm2 and the coercive electric field [Ec] is 5.4 kV/cm for 0.98NKN-0.02BZT ceramics sintered at 1,080°C. When the sintering temperature was increased up to 1,090°C, the well-saturated ferroelectric properties were obtained, and the values of Pr and Ec of samples sintered at 1,090°C were 10.3 μC/cm2 and 7.2 kV/cm, respectively. Continuously increasing the sintering temperature above 1,090°C, the ferroelectric properties decreased due to heavy volatilization of Na and K at high sintering temperature. The ferroelectric properties of 0.98NKN-0.02BZT ceramics have a similar tendency as with the piezoelectric and dielectric properties. The increase of ferroelectric properties might be caused by the increase of the relative bulk density that reduces the leakage current, promoting the polarization process.where d33 is 217 pC/N, which is the highest value in the 0.98NKN-0.02BZT system. The equivalent properties of Tc, kp, Pr, and dielectric constant values are 411°C, 0.41, 10.3 μC/cm2, and 1,951, respectively. Therefore, 0.98NKN-0.02BZT ceramics is a potential candidate for lead-free piezoelectric ceramics.In conclusion, the lead-free 0.98NKN-0.02BZT ceramics with a perovskite structure have been sintered at various sintering temperatures. The effects of the sintering temperatures on the structural and electrical properties were investigated. Increasing sintering temperatures improve the grain growth, densification, and electrical properties in effect. However, with an increasing sintering temperature above 1,090°C, the structural and electrical properties have significantly deteriorated. The obtained The authors declare that they have no competing interests.S-HL carried out the experiments which show the electrical properties and drafted the manuscript. S-GL carried out the experiments which show the structural properties and reviewed the manuscript. Y-HL participated in the design of this study and reviewed the manuscript finally. All authors read and approved the final manuscript.


Smenospongia aurea led to the isolation of smenamide A (1) and B (2), hybrid peptide/polyketide compounds containing a dolapyrrolidinone unit. Their structures were elucidated using high-resolution ESI-MS/MS and homo- and heteronuclear 2D NMR experiments. Structures of smenamides suggested that they are products of the cyanobacterial metabolism, and 16S rRNA metagenomic analysis detected Synechococcus spongiarum as the only cyanobacterium present in S. aurea. Smenamides showed potent cytotoxic activity at nanomolar levels on lung cancer Calu-1 cells, which for compound 1 is exerted through a clear pro-apoptotic mechanism. This makes smenamides promising leads for antitumor drug design.An in-depth study of the secondary metabolites contained in the Caribbean sponge  Smenospongia aurea , collected by Scuba along the coast of Little Inagua (Bahamas Islands).As part of a broad screening program going on at the NeaNat research group focused on the study of sponge natural product chemistry in order to discover novel lead molecules for anticancer  and antiSmenospongia, are a well-known sources of many interesting secondary metabolites. Among them, it is worth mentioning the large number of indole alkaloids +) in the HRMS/MS spectrum confirmed the presence of chlorine in the molecule.The positive ion mode high-resolution ESI mass spectrum of smenamide A (1)  displaye1H NMR spectrum of compound 1 (3-27) and the methine singlets at δ 5.93 and 5.97 (H-21)  suggested smenamide A (1) to be the product of a PKS/NRPS pathway.The molecular formula and the general features of the N-methyl group  and the acetyl methyl group  showed correlation peaks in the HMBC spectrum with the same carbonyl carbon atom at δ 172.9, suggesting an N-methylacetamido function. This was confirmed by the long-range coupling between the two methyls through the amide bond + ion at and 523.2320). The general features of the 1H NMR spectrum (Z in smenamide B on the basis of the correlation peak between H3-14 (δ 1.79) and H-15 (δ 5.13) observed in the ROESY spectrum.Smenamide B . TherefS. aurea collected in different years and geographical areas, suggesting their symbiotic, rather than dietary, origin.The structural motifs of smenamide A and B strongly suggest that they are a product of the cyanobacterial metabolism. In addition, smenamides have been found in comparable amounts in three different specimens of S. aurea was examined by biomolecular analysis. A PCR experiment performed using the metagenome of S. aurea as template and the cyanobacterial-specific 16S rRNA primers CYA106F and CYA781R(a) or CYA781R(b) , thus affording a fraction (Rt = 10 min) containing compound 1 and a fraction (Rt = 8 min) containing the compound 2. The former fraction was subjected to normal-phase HPLC on an SiO2 column , which gave 15 µg of pure compound 1.The total organic extract (12.25 g) was chromatographed on a column packed with RP-18 silica gel. A fraction eluted with MeOH/H2 was first subjected to HPLC in the same condition as for 1; final purification was achieved by HPLC on an analytical RP-18 column using CH3CN/H2O as eluent , to obtain pure compound 2 (8 µg).The fraction containing compound S. aurea, collected in 2007 at Sweeting Cay (Grand Bahama Island) and in 2008 at Little San Salvador (Bahamas Islands), respectively, showed them to contain similar amounts of compounds 1 and 2.LC-MS analysis of the crude extract of two additional specimen of m/z 523.2326 , m/z 501.2509 ; MS isotope pattern: M (100%), M + 1 , M + 2 , M + 3 ; HRESIMS/MS : m/z 487.2557 , 397.2092 , 320.1384 , 284.1618 , 244.0941 , 226.0836 , 202.0472 ; 1H and 13C NMR: max(ε) 287 nm (8200), 246 nm (46000), 225 nm (92000); CD (MeOH): λmax(Δε) 238 (+33), 219 (−30).Colorless amorphous solid, HRESIMS  m/z 523.2320 , m/z 501.2505 ; MS isotope pattern: M (100%), M + 1 , M + 2 , M + 3 ; HRESIMS/MS : m/z 487.25567 , 397.2091 , 320.1383 , 284.1617 , 244.0941 , 226.0835 , 202.0471 ; 1H and 13C NMR: 1H and 13C NMR: max(ε) 287 nm (15000), 248 nm (42000), 225 nm (84000); CD (MeOH): λmax(Δε) 237 (+7.3).Colorless amorphous solid, HRESIMS  ®-H2O) and 50 µL lysozime (20 mg/mL) were added and incubated at 37 °C for 1 h in a thermomixer (1400 rpm). After addition of 2.8 µL β-mercaptoethanol, 70 µL 10% SDS, 2 µL RNase A (100 mg/mL), and 40 µL proteinase K (10 mg/mL) the tube was incubated at 55 °C for a further hour in a thermomixer (1400 rpm). At this time, the microcentrifuge tube was spun 4 min at 5000 rpm. The clear middle phase was transferred to a new microcentrifuge tube containing 750 µL CHCl3 and centrifuged 10 min at 15,000 rpm. After two further CHCl3 washes, the supernatant was transferred to a new microcentrifuge tube containing 750 µL of 70% aqueous isopropyl alcohol containing 10% (v/v) 3 M NaOAc (pH 5.5) at room temperature. The precipitated DNA was spun down at top speed for 20 min (4 °C), washed with ice-cold ethanol, dried and dissolved in ~100 µL of water.To ~40 mg of frozen sponge (in RNA later) 700 µL of lysis buffer I , 2.5 µL Taq buffer advanced , 2.5 µL forward primer CYA106F (10 µM), 2.5 µL reverse primer CYA781R(a) or CYA781R(b) (10 µM), 0.35 µL RBC Taq DNA polymerase , 0.5 µL DNA. The PCR products of the expected size (~670 bp) were purified from the agarose gel using QIAquick gel ex kit , subcloned via T/A cloning into pBluescriptII SK(+) , and transferred to E. coli XL1 Blue MRF'  electrocompetent cells. After blue white screening, plasmid preps were sent to GATC Biotech AG  for single read sequencing using the T7 primer.Metagenomic DNA from S. aurea was used for PCR amplification with the 16S rRNA cyanobacterial specific primers CYA106F (5′-CGGACGGGTGAGTAACGCGTGA-3′), CYA781R(a) (5′-GACTACTGGGGTATCTAATCCCATT-3′) and CYA781R(b) (5′-GACTACAGGGGTATCTAATCCCTTT-3′). Polymerase chain reaction was carried out under the following conditions: initial denaturation at 94 °C for 45 s, followed by 33 cycles of 94 °C for 1 min, 60 °C/64 °C for 1 min and 72 °C for 45 s, with a final extension 72 °C for 10 min. The reaction mixture (25 µL) contained: 14.65 µL H2 atmosphere. Cells were harvested at 80% confluence to perform further examination.Calu-1 cell line was purchased from the American Type Culture Collection (ATCC). Cell line was grown in Iscove’s modified Dulbecco’s medium (IMDM) , supplemented with 10% FBS , 100 units/mL Penicillin plus 100 µg/mL Streptomycin , 250 mg/mL Amphotericine B . Cell cultures were incubated at 37 °C in a humidified 5% CO3 cells/well, in a total volume of 100 μL of complete medium. After treatment for 72 h with compounds 1 and 2 at different concentrations , MTT (5 mg/mL in PBS) was added to each well, and the cells were incubated for 4 h at 37 °C. Dimethylsulfoxide (DMSO) was added to each well after removal of the supernatant, and the plates were shaken to dissolve formazan. The absorbance reading of each well was determined using a computer-controlled microtiter plate reader  at a wavelength of 540 nm. All experiments were performed in triplicate for each condition.Cell viability was determined by 3--2,5-diphenyltetrazolium bromide (MTT) assay according to the manufacturer’s instructions. Briefly, cells were plated in 96-well plates, at a density of 3 × 10p < 0.05. The IC50 values were determined by nonlinear regression analysis using a 4-parameter logistic equation.The optical density of the cells incubated with culture medium alone was taken as 100% viability. Differences between groups were determined by analysis of variance (ANOVA) and were considered statistically significant at 5 cells/well and were treated for 72 h with compounds 1 and 2 at concentrations 1, 10, 50, and 100 nM. After treatment cells were recovered, washed with Binding Buffer (BB) and centrifuged. Annexin-V FITC (10 μL) was added to the pellets, which were then incubated in the dark for 15 min at room temperature. Next, cells were washed and re-suspended in BB, and propidium iodide solution (5 μL) was added immediately prior to analysis by flow citometry. The percentage of apoptosis was analyzed by Diva Software using a Becton Dickinson FACS ARIA II.Apoptosis was assessed using the Annexin-V FITC kit  according to manufacturer’s instructions. Briefly, cells were plated in 6-well plates at a density of 100 × 10The cyanobacterial 16S rRNA partial sequences from metagenomic analysis were deposited into GenBank under the accession numbers KF692286 through KF692298 (primers CYA106F/CYA781R(a)), and KF692299 through KF692309 (primers CYA106F/CYA781R(b)).The discovery of smenamides further expands the catalog of highly bioactive marine natural products, and can potentially provide new templates for the development of antitumor drugs.1) and B (2) are two isomeric hybrid peptide/polyketide compounds containing a dolapyrrolidinone unit, which are present in low concentrations in the marine sponge S. aurea. In spite of the very little amounts of the isolated compounds (15 μg for 1 and 8 μg for 2), their planar structures were completely determined by spectroscopic means.Smenamide A (S. aurea belong to Synechococcus spongiarum, a symbiotic cyanobacterium widespread in tropical and temperate reef sponges.The structural features of smenamides suggest them to be products of cyanobacterial metabolism; the only cyanobaterial 16S rRNA sequences detected in the metagenome of 1. Therefore, while displaying similar IC50 inhibitory values for human Calu-1 lung cancer cells, compounds 1 and 2 do not seem to exert their respective activity through the same mechanism. Further studies are in progress to gain a better insight into the antitumor activity of smenamides. It is important to point out that the widespread diffusion of S. aurea will make it possible to isolate higher amounts of smenamides, making them amenable for further development as antitumor compounds.Smenamides showed potent cytotoxic activity at nanomolar level on lung cancer Calu-1 cells, which is exerted through a clear pro-apoptotic mechanism only for compound


C1, C2 and C3 in acyclic diene metathesis (ADMET) polymerization of a fully renewable α,ω-diene. A high activity of these catalysts was observed for the synthesis of the desired renewable polyesters with molecular weights of up to 17000 Da, which is considerably higher than molecular weights obtained using the same monomer with previously studied catalysts. Moreover, olefin isomerization side reactions that occur during the ADMET polymerizations were studied in detail. The isomerization reactions were investigated by degradation of the prepared polyesters via transesterification with methanol, yielding diesters. These diesters, representing the repeat units of the polyesters, were then quantified by GC-MS.We report on the catalytic activity of commercially available Ru-indenylidene and “boomerang” complexes  Among the large number of organic and organometallic reactions allowing the formation of carbon–carbon bonds, olefin metathesis has found its place in organic synthesis as well as polymer science as a very versatile tool that allows transformations that were previously not (or hardly) possible –6. This st generation catalyst was capable of catalyzing olefin isomerization [CF3SO3], as the catalyst precursor and HOSO2CF3, and applied in the ADMET of 1,9-decadiene to yield a polymer with 94% conversion in 12 h at 0 °C.To date, only one example of ADMET polymerization with an in situ generated Ru–indenylidene catalyst has been reported . The relC1 and C2, and the “boomerang complex” C3 , under bulk conditions, after 5 h reaction time, and constant catalyst loading . This provided a broad data set to screen the catalytic systems tested , which was previously examined in ADMET polymerizations of the same monomer methylene ruthenium(II) , -2-imidazolidinylidene)dichloro(o-isopropoxyphenylmethylene) ruthenium(II) .10-undecenoic acid , 1,3-propanediol , Thin layer chromatography (TLC) was performed on silica gel TLC-cards . Compounds were visualized by permanganate reagent. For column chromatography silica gel 60  was used.1H NMR spectra were recorded in CDCl3 on Bruker AVANCE DPX spectrometers operating at 300 and 500 MHz. Chemical shifts (δ) are reported in parts per million relative to the internal standard tetramethylsilane . For the analysis of the polymers the relaxation time was set to 5 seconds.TM option installed. Samples were introduced by direct infusion with a syringe pump. Nitrogen served both as the nebulizer gas and the drying gas. Helium was used as cooling gas for the ion trap and collision gas for MSn. Nitrogen was generated by a nitrogen generator Nitrox from Dominick Hunter.Mass spectra (ESI) were recorded on a VARIAN 500-MS ion trap mass spectrometer with the TurboDDS−1. The oven temperature was programmed as follows: initial temperature 95 °C, hold for 1 min, ramp at 15 °C × min−1 to 200 °C, hold for 2 min, ramp at 15 °C × min−1 to 325 °C, hold for 5 min. The injector transfer line temperature was set to 250 °C. Measurements were performed in the split–split mode (split ratio 50:1) with helium as carrier gas (flow rate 1.0 ml × min−1).GC-MS (EI) chromatograms were recorded with a Varian 431-GC instrument with a capillary column FactorFourTM VF-5ms (30 m × 0.25 mm × 0.25 μm) and a Varian 210-MS detector. Scans were performed from 40 to 650 m/z at a rate of 1.0 scans × s−1) at 50 °C. All determinations of molar mass were performed relative to PMMA standards .Polymer molecular weights were determined with an SEC System LC-20 A from Shimadzu equipped with a SIL-20A auto sampler, three PSS SDV columns , and a RID-10A refractive index detector in THF (ﬂow rate 1 mL × min−1 with a DSC821e (Mettler Toledo) calorimeter up to a temperature of 150 °C with a sample mass of approximately 4 mg. The melting temperature, Tm, was recorded as the peak of the endotherm on the second heating scan unless annealing was used as a pretreatment.Differential scanning calorimetry (DSC) experiments were carried out under a nitrogen atmosphere at a heating rate of 10 °C × minp-toluensulfonic acid  were placed in a round-bottomed flask provided with a magnetic stirrer and a Dean-Stark apparatus. Toluene (200 mL ) was added and the resulting reaction mixture heated to reflux. Water was collected as the reaction proceeded and once the reaction was completed, the reaction mixture was allowed to cool. Toluene was removed under reduced pressure and the residue was filtered through a short pad of basic aluminium oxide with hexane as eluent. After removing the hexane, the crude product was dissolved in diethyl ether (200 mL) and washed two times with water (200 mL). The organic fraction was dried over anhydrous MgSO4 and the solvent removed under reduced pressure. The desired product was isolated in 87% yield (39 g).10-Undecenoic acid , 1,3-propanediol  and 1H NMR (CDCl3): δ = 5.85–5.76 , 5.00–4.91 , 4.15 , 2.30 , 2.00 , 1.99–1.94 , 1.64–1.58 , 1.38–1.34  1.29–1.24  ppm. 13C NMR (CDCl3): δ = 173.6 , 139.0 , 114.1 , 60.7 , 34.1 , 33.7 , 29.2 , 29.1 , 29.0 , 28.8 , 24.8  ppm. MS (EI): m/z = 408 [M]+, calc. 408.3239.C1: 11.6 mg (0.0122 mmol), C2: 9.1 mg (0.0122 mmol), C3: 8 mg (0.0122 mmol) and C4: 7.7 mg) was added at the desired reaction temperature (60–120 °C). In some cases, 1 mol % of BQ was added to the reaction mixture 10 min before the addition of the catalyst. Reactions were carried out in parallel using a carousel reaction station from Radleys. Stirring was continued at the selected temperature under a continuous flow of nitrogen for 5 h. After 5 h reaction time, the reaction mixture was dissolved in 1 mL of THF and polymerization halted by the addition of 1 mL of ethyl vinyl ether. The mixture was then stirred for 30 min at room temperature. The crude product was purified by precipitation into cold methanol. Final polymer molecular weights were determined after precipitation with the above mentioned GPC system.To 1 g (2.45 mmol) of 1,3-propylene diundec-10-enoate in a tube equipped with a screw, 0.5 mol % of the corresponding ruthenium catalyst, (The respective polymer (30 mg), excess methanol (4 mL) and concentrated sulfuric acid (5 drops) were added to a carousel reaction tube, stirred magnetically, and refluxed at 85 °C for 5 h. At the end of the reaction, the excess of methanol was removed under reduced pressure. The residue was then dissolved in diethyl ether and filtered through a small column of basic aluminium oxide. Subsequently, GC-MS samples were prepared by taking 500 µL of this solution and diluting it with methanol (500 µL). The percentage of olefin isomerization was calculated based on peak areas of the isomerized diesters.


There are several advantages of using the LMO/G as cathodes in Li-ion batteries: (1) the LMO/G electrode shows high specific capacities at high gravimetric current densities with excellent cycling stability, e.g., 84 mAh·g−1 during the 500th cycle at a discharge current density of 5625 mA·g−1 (~38.01 C capacity rating) in the voltage window of 3–4.5 V; (2) the LMO/G hybrid can buffer the Jahn–Teller effect, which depicts excellent Li storage properties at high current densities within a wider voltage window of 2–4.5 V, e.g., 93 mAh·g−1 during the 300th cycle at a discharge current density of 5625 mA·g−1 (~38.01 C). The wider operation voltage window can lead to increased theoretical capacity, e.g., 148 mAh·g−1 between 3 and 4.5 V and 296 mAh·g−1 between 2 and 4.5 V; (3) more importantly, it is found that the attachment of LMO onto graphene can help to reduce the dissolution of Mn2+ into the electrolyte, as indicated by the inductively coupled plasma (ICP) measurements, and which is mainly attributed to the large specific surface area of the graphene sheets.We report a facile method to prepare a nanoarchitectured lithium manganate/graphene (LMO/G) hybrid as a positive electrode for Li-ion batteries. The Mn Spinel Lithium Manganate (LMO) is a promising cathode material for rechargeable Li-ion batteries due to the high energy density, improved operating safety, low cost and low toxicity [−6 S·cm−1) and poor cycling performance of LMO are the main issues associated with this material [2+ (formed through Mn3+ disproportionation) dissolution into the electrolyte, (2) the electrolyte decomposition and (3) the Jahn–Teller effect [Lithium-ion batteries (LIBs) are considered the primary candidate as the power source for plug-in and hybrid electric vehicles . Althougmaterial –4. The pr effect , which r3+ in LMO, e.g., by replacing manganese with monovalent [−1 at 1 C rate (capacity rating) [In order to solve these issues, many methods have been implemented. In general, these methods can be divided into two categories: chemical doping and microstructure modification. The chemical doping method involves elevating the average manganese ion oxidation state so as to decrease the amount of Mnnovalent  or multinovalent –9. Howevnovalent –13. Orde rating) . The sil rating) 15–16 can2S into the electrolyte during cycling. This could be considered as a useful strategy to improve the dissolution of Mn2+ from the LMO cathode. However, the preparation of graphene-based cathodes is still limited [The fabrication of nanoarchitectured 3D electrodes is a promising approach to improve the power density and cyclability of LIBs –18. Basi limited ,28. A poCurrently, the reported synthesis of graphene-based electrode materials for LIB is mostly based on the method of Hummers –2629–32,262O3 nanowalls in a one-step electrochemical process, followed by molten salt lithiation to convert Mn2O3 to LMO. The molten salt process [4 in the electrochemical process. The LMO/G hybrid cathode shows excellent Li storage properties under fast lithium intercalation/deintercalation processes. The optimized cathode delivers a stable specific capacity of ~84 mAh·g−1 during the 500th cycle at a gravimetric current density of 5625 mA·g−1 (~38.01 C) with a voltage window of 3–4.5 V, which corresponds to a specific power density of 100 kW·kg−1 and a specific energy density of 278 Wh·kg−1. Also, the LMO/G hybrid can buffer the Jahn–Teller effect [−1 during the 300th cycle at a discharge current density of 5625 mA·g−1 (~38.01 C). Last and most important, it is found that the dissolution of Mn2+ into the electrolyte is very much reduced for the LMO/G sample as compared to that of commercially purchased LMO, as indicated by the inductively coupled plasma (ICP) study. This is important for achieving a stable performance of the LMO-based cathodes. The methodology proposed here should also be applicable for the preparation of other lithium-metal-oxide/graphene hybrids for high-power LIB applications.Electrochemical exfoliation is a newly developed method that can be used to prepare highly conductive graphene sheets in high yield and at low cost . Moreove process –37 requi4 using graphite sheets as the working electrodes. In this process, metal-oxide deposition and graphene exfoliation are implemented simultaneously. The XRD patterns (see 2O3 (JCPDS No. 41-1442) phase in the various samples prepared, by changing the concentration of MnSO4. The grain sizes of Mn2O3 in the various samples are estimated to be about 20 nm, based on the full width at half maximum of the diffraction peaks by using Scherrer’s equation. The bump at around 2θ = 25° is attributed to the glass holder. There are no other detectable peaks corresponding to impurity phases in the XRD patterns.The samples from the electrochemical deposition were prepared by the electrolysis of MnSOerns see  confirm 4 molar concentration of 0.15 M, the SEM image (see 2O3 phase. The observed interlattice spacing of 0.470 nm in the HRTEM corresponds to the (200) planes of the cubic Mn2O3 (JCPDS No. 41-1442) phase.The SEM and TEM images of the as-prepared samples are shown in mage see  shows thmage see  reveals mage see  and the mage see  confirms2D/G of the 2D band (located at 2720 cm−1) to the G band (located at 1580 cm−1) is 0.16, which is similar to that reported for electrochemically exfoliated graphene [MO:G, of Mn2O3:graphene are found to be directly related to the concentration of MnSO4 in the precursors. The thermal gravimetric analysis (TGA) results . Here, the determination of the exact phase of the sample after the lithiation and annealing process is based on both the XRD analysis and inductively coupled plasma (ICP) measurements. The XRD pattern of cubic spinel LiMn2O4 (JCPDS No. 35-0782) is similar to that of cubic spinel Li4Mn5O12 (JCPDS No. 046-0810). The ICP results  after the lithiation and annealing process. These ICP results are approximately the ideal ratios for LiMn2O4, which confirms the stoichiometry of the final samples. The grain size of the spinel LiMn2O4 (LMO) is estimated to be only 3.2 nm, based on the full width at half maximum of the diffraction peaks using Scherrer’s equation. This grain size is much smaller than that of Mn2O3, which is possibly due to a recrystallization process. No impurity phase is detectable using XRD.The as-prepared Mnreaction  under Arerns see  of the sults see  indicateLMO:G, between the LMO and graphene are 1.22, 2.00 and 4.49 for the samples prepared by lithiating Mn2O3/G samples with IMO:G = 0.82, 1.34 and 2.57, respectively. Here, the calculated ILMO:G values are 1.07, 1.90, and 4.71 for Mn2O3/G samples with IMnO:G values of 0.82, 1.34 and 2.57, respectively, which are close to the measured ILMO:G values.After the lithiation process, the TGA results  and G band increases after the lithiation process, which indicates the increased number of defects in the graphene sheets.The Raman spectra show that the intensity ratio of the 2D band to the G band , which is consistent with the XRD results. The HRTEM image  planes of the cubic spinel LiMn2O4 phase (JCPDS No. 35-0782).For the LMO/G sample with Image see  shows thtion see . The SAEtion see  obtainedmage see  shows thLMO:G values, the SEM images , for the second cycle at 1.27 C rate (187.5 mA·g−1) between 3 and 4.5 V. The 4 V plateau during discharge corresponds to the reduction of Mn4+ to Mn3.5+ through the reaction of Li1−xMn2O4 + xe− + xLi+ → LiMn2O4, where x is ~1 [To evaluate the cathode performance of the LMO/G hybrids, half cells were fabricated based on a Swagelok configuration –40. Figu x is ~1 . The hig x is ~1 .LMO:G = 1.22) electrode is evaluated under different discharge currents between 3 and 4.5 V. LMO:G = 1.22) electrode delivered second-cycle discharge capacities of 108, 101, 94.5, 87, 81, 76.5, 65.5 and 51.5 mAh·g−1 at current densities of 187.5 (1.27 C), 375 (2.53 C), 937.5 (6.33 C), 1875 (12.67 C), 3750 (25.34 C), 5625 (38.01 C), 9375 (63.34 C) and 18750 (126.68 C) mA·g−1, respectively. These values correspond to a series of energy densities of 402, 378, 353, 323, 299, 278, 236 and 175 Wh·kg−1 at different power densities of 3.37, 6.80, 17.17, 34.20, 67.28, 100.08, 163.39 and 315.00 kW·kg−1, respectively. The LMO/G electrodes (ILMO:G = 1.22) present higher energy densities at high power densities than porous LMO (282 Wh·kg−1 at 15 kW·kg−1) [−1 at 10 kW·kg−1) [−1 at 54 kW·kg−1) and LMO nanotubes (304 Wh·kg−1 at 5.6 kW·kg−1) [The rate capability of the LMO/G (IkW·kg−1) , porous kW·kg−1) , single kW·kg−1)  (243 Wh·kW·kg−1) .−1 at current densities of 187.5 (1.27 C), 375 (2.53 C), 937.5 (6.33 C), 1875 (12.67 C), 3750 (25.34 C), 5625 (38.01 C), 9375 (63.34 C) and 18750 (126.68 C) mA·g−1, respectively. These values correspond to a series of energy densities of 382, 310, 282, 254, 226, 194, 155 and 117 Wh·kg−1 at different power densities of 3.17, 6.36, 16.07, 32.69, 62.05, 85.59, 104.57 and 134.72 kW·kg−1, respectively. The commercially purchased pure LMO powder shows comparable discharge capacities to that of LMO/G electrodes at low current densities, e.g., <3 C. However, at high current densities, e.g., >25 C, the LMO/G (ILMO:G = 1.22) electrodes show much higher discharge capacities than that of pure LMO, which indicates that the conducting scaffold of graphene sheets improves the kinetics of Li ion diffusion and electron transfer.For the purpose of comparison, the rate capability of commercially purchased pure LMO powders was also investigated between 3 and 4.5 V see . The corLMO:G = 1.22) electrode under various discharge currents are presented in −1.The voltage profiles of the LMO/G (ILMO:G = 1.22) electrode is evaluated (see −1 (1.27 C), 937.5 mA·g−1 (6.33 C) and 5625 mA·g−1 (38.01 C) in the voltage range of 3–4.5 V. When discharging at 1.27 C and 6.33 C, the LMO/G (ILMO:G = 1.22) electrode delivered discharge capacities of 110 mAh·g−1 and 100 mAh·g−1, respectively, during the second cycle, which did not show any obvious decrease and remained at 110 mAh·g−1 and 100 mAh·g−1, respectively, during the 500th cycle with capacity retentions of 100% per cycle. Even at a very high discharging rate of 38.01 C, it exhibited highly reversible specific capacities of 79 mAh·g−1 during the second cycle and 84 mAh·g−1 during the 500th cycle. The cycling performance of LMO/G with ILMO:G = 2.00 and 4.49 are also evaluated at 1 C in the voltage range of 3–4.5 V  for the second cycle at 1.27 C rate (187.5 mA·g−1) between 2 and 4.5 V. The plateau at ~3 V corresponds to the further reduction of Mn3.5+ to Mn3+ through LiMn2O4 + ye− + yLi+ → Li1+yMn2O4, where y is ~1 [It is known that the spinel LMO phase undergoes an additional redox reaction at 3 V, which increases the theoretical capacity, e.g., 148 mAh·gl spinel , which r y is ~1 . The symLMO:G = 1.22) electrode was evaluated under the same charge/discharge currents between 2 to 4.5 V. LMO:G = 1.22) electrode delivers second-cycle discharge capacities of 245, 238, 217, 196, 167 and 148 mAh·g−1 at current densities of 187.5 (1.27 C), 375 (2.53 C), 937.5 (6.33 C), 1875 (12.67 C), 3750 (25.34 C) and 5625 mA·g−1 (38.01 C), respectively. After cycling at 5625 mA·g−1 (38.01 C), the discharge capacity of the LMO/G (ILMO:G = 1.22) electrode recovers back to 257 mAh·g−1 when changing the current density to 187.5 mA·g−1 (1.27 C), showing a good rate performance. The high C-rate performance is slightly worse for samples with higher ILMO:G values (see −1 (38.01 C), the LMO/G (ILMO:G = 2.00) and LMO/G (ILMO:G = 4.49) electrodes deliver discharge capacities of 128 and 117 mAh·g−1, respectively, during the second cycle.The rate capability of the LMO/G  electrode was also evaluated at very high current densities, e.g., a discharge current density of 5625 mA·g−1 (38.01 C) and a charge current density of 2812.5 mA·g−1 (19.00 C), between 2 and 4.5 V (see LMO:G = 1.22) electrode depicts a specific capacity of 170 mAh·g−1 during the second cycle, which remains at 93 mAh·g−1 during the 300th cycle. Based on the voltage profiles under such cycling conditions, the corresponding energy density and power density were calculated to be 284 Wh·kg−1 and 17.04 kW·kg−1, respectively, during the 300th cycle. Here, it is worth pointing out that the charge rate is chosen as 19.00 C (2812.5 mA·g−1) to mimic the fast battery charging process. In fact, some reports on the rate capabilities of cathode materials use a high discharge C rate and a low charge C rate, which are different from the really fast battery charging process, especially for plug-in electric vehicles. For LMO/G electrodes with ILMO:G = 2.00 and 4.49, the cycling responses under similar testing parameters, e.g., a discharge current density of 5625 mA·g−1 (38.01 C) and a charge current density of 2812.5 mA·g−1 (19.00 C), are also evaluated  at 50 °C for 72 h. The manganese concentrations in the electrolytes are 11.76, 14.22, 17.82 and 23.35 ppm . This is possibly due to the wrapping of graphene sheets onto the LMO nanocrystals and the very large specific surface area of the graphene sheets, e.g., >100 m2·g−1. Thus, the graphene sheets may act as a physical adsorption layer to anchor the Mn2+ ions. Increasing the graphene content in the hybrid sample helps to reduce the dissolution of Mn into the electrolyte further.For LMO cathode, there is an additional concern about the dissolution of the Mnctrolyte . Thus, t ppm see  for LMO/2+ cations and thereby enhance the cyclability of the electrodes.The superior electrochemical performance of LMO/G electrodes is ascribed to three aspects. First, the LMO/G exhibits fast kinetics of Li-ion and electron diffusion, as examined by the electrochemical impedance spectroscopy, which suggests that the 3D conducting scaffold formed by graphene sheets and the fine grain size of the LMO achieved by such a synthesis process help to improve the Li-storage performance of the hybrid electrodes at high current densities. Second, the LMO/G electrodes are demonstrated to show improved capacity retention at high current densities in the voltage range of 2–4.5 V. For the LMO/G hybrids, the fine LMO grains  are embedded in the 3D conducting scaffold formed by the graphene sheets, which can effectively relieve the issue caused by the Jahn–Teller effect, by allowing the LMO grains to expand/contract freely with the support of the conducting graphene sheets. Finally, the incorporation of graphene scaffold is herein proved to be an effective way to suppress Mn dissolution into the electrolyte. In fact, Mn dissolution is one of the biggest concerns for LMO, which significantly affects the cycling stability. It is especially serious for nanosized LMO due to their large surface area exposed to the electrolyte. Mn dissolution causes the collapse of the crystal structure, which leads to the capacity losses during cycling. For LMO/G hybrids, the graphene sheets serve as a physical adsorption layer of anchoring Mn2O3 and exfoliation of graphite electrodes in a one-step electrochemical process, followed by molten salt lithiation. The weight ratios between the LMO and graphene can be readily adjusted by simply changing the concentration of Mn2+ in the electrochemical process. These LMO/G electrodes show excellent cathode performances with high specific capacities and stable cyclability at high current densities in different voltage ranges, e.g., 2–4.5 V and 3–4.5 V. This is mainly due to the several advantages of such hybrids: (1) The combination of the highly conductive 3D graphene scaffold and the small grain size of the LMO  greatly enhance the kinetics of charge transfer in the electrode; (2) the flexible graphene sheets can buffer the volume strain of the LMO grains caused by the transition from cubic to tetragonal spinel and, hence, improve the cycling stability in the voltage range of 2–4.5 V; and (3) the graphene sheets may effectively reduce the dissolution of Mn into the electrolyte and improve the cycling performance.In conclusion, a novel and facile approach was developed to synthesize a nanoarchitectured LMO/G hybrid as a positive electrode material for lithium-ion batteries. The process combines the deposition of Mn2O3 and graphene hybrids were prepared by electrochemical method. Highly oriented pyrolytic graphite sheets  and platinum wires (0.2 mm in diameter) were used as the working electrodes and counter electrodes, respectively. The electrolyte was prepared by mixing MnSO4, CH3COONa and Li2SO4 with deionized water. The concentrations of Mn2+ were selected to be 0.15, 0.3 and 0.6 M. The molar concentrations of CH3COONa were equivalent to that of MnSO4. The concentrations of Li2SO4 were 0.85, 0.7 and 0.4 M for the three solutions with 0.15, 0.3 and 0.6 M MnSO4. The electrochemical cells were heated to 60 °C and a stepwise voltage of 0–10 V was applied between the working electrode and the counter electrode for 30 min. The products were collected through vacuum filtration, washed several times with deionized water, and centrifuged to remove large graphite particles, followed by drying at 50 °C overnight.Mn3 was dissolved in deionized water with the addition of 50 mg of as-prepared Mn2O3/graphene. After vigorous stirring, the solution was dried at 80 °C and then heated to 300 °C in argon gas for 2 h. The resultants were rinsed with deionized water. Thereafter, the final products were annealed at 350 °C for 30 min in argon gas.The lithiation process was carried out via molten salt reaction. A mixture of 9.9 mg LiNO−1 in air. The analyses of Li/Mn ratio was performed on inductively coupled plasma atomic emission spectrometer .Powder XRD patterns were obtained on a Shimadzu XRD-6000 X-ray diffractometer with Cu Kα irradiation. SEM imaging was performed on a JEOL JSM-7600F operating at 5 kV. TEM and HR-TEM images were obtained on a JEOL JEM-2100 operating at 200 kV. Raman spectra were obtained on a WITec CRM 200 with a wavelength of 488 nm and a spot size of 0.5 mm. Thermogravimetry analyses were carried out on TGA Q500 from 298–1073 K at a heating rate of 10 K·min6 in EC/DEC at 50 °C for 72 h. The organic solvent was evaporated and the sediments were dissolved in aqueous 4 M HNO3, followed by examination of the manganese concentrations with ICP-AES (PerkinElmer OPTIMA 2000 DV). The results were calculated back to the original manganese concentrations in the electrolyte.Mn dissolution was examined by immersing LMO/G and pure LMO with identical weight of 100 mg LMO in 5 mL LiPFN-methyl-2-pyrrolidone. The mixture was stirred overnight and then cast onto aluminium foils to form uniform electrodes followed by drying in vacuum at 50 °C for 10 h. As for pure LMO electrodes, commercial LMO , Super-P (10 wt %) and poly(vinylidene fluoride) (10 wt %) were mixed and cast onto aluminium foils. CR2032 coin cells were assembled in an argon-filled glove box with the contents of oxygen and moisture below 0.1 ppm, using lithium foils as the counter and reference electrode and 1 M LiPF6 in ethylene carbonate and dimethyl carbonate (1:1 v/v) as the electrolyte. All cells were tested on a NEWARE system at 4.5–3 V versus Li+/Li. All capacities were calculated based on the mass of LMO and all C-rates were calculated according to the theoretical capacity of 148 mAh·g−1. The electrochemical impedance was performed on the aforementioned CR2032 coin cells with lithium foil as the counter and reference electrode. All cells were measured at the fifth fully discharged state. INPHAZETM EIS system was employed to measure the electrochemical impedance spectra. The amplitude of the alternating voltage was set at 5 mV and the frequency range was 1000 kHz to 10 MHz.Electrochemical measurements were carried out on half cells. The electrodes were fabricated by mixing LMO/G (90 wt %) and poly(vinylidene fluoride) (10 wt %) in Raman spectrum, TGA results, SEM and HRTEM images and electrochemical performance figures.File 1Additional figures.


Self-help or self-management strategies are commonly used to deal with depression, but not all are thought to be helpful. A previous study found that sub-threshold depression symptoms were improved by an e-mail intervention that encouraged the use of evidence-based self-help strategies.To investigate whether these e-mails were effective for adults with a range of depression symptomatology including major depression.The study was a parallel-group randomised controlled trial. Adult participants with any level of depressive symptoms were recruited over the internet from the United Kingdom, Australia, Canada, Ireland, New Zealand and the United States. Participants were randomised to receive a series of e-mails either promoting the use of evidence-based self-help strategies or containing depression information as a control. E-mails were sent automatically twice a week for six weeks. Depression symptoms were assessed with the self-rated Patient Health Questionnaire depression scale (PHQ-9).d = 0.11, 95% CI: −0.06 to 0.27), although there was a small effect at the study mid-point. Results were similar for the sub-group of participants with major depression. The active group showed small to moderate improvements in self-help behaviour .1736 participants with a wide range of symptom severity were recruited and assigned to active (n = 862) and control (n = 874) groups. However, there was a significant attrition rate, with 66.9% lost to follow-up at post-intervention. Both groups showed large improvements in depression symptoms overall, with no significant difference in improvement at the end of the study (mean difference in improvement 0.35 points, 95% CI: −0.57 to 1.28, These results suggest that the e-mails were able to increase participants’ use of evidence-based self-help, but that this did not improve depression more than an attention control.NCT01399502 Geisner, Neighbors and Larimer To investigate whether e-mails promoting effective self-help strategies could change self-help behaviour and improve depression, we developed an e-mail-based intervention called Mood Memos. This was a fully automated system that could be easily disseminated. It delivers 12 e-mails over a 6-week period, which encourage the use of self-help strategies endorsed by experts for depression. A randomised controlled trial indicated that the e-mails were effective for improving sub-threshold levels of depression Given this promising result, we were interested in whether the Mood Memos intervention would be helpful for those with full-threshold levels of depression, as the first study targeted only mild or sub-threshold depressive symptoms. Evaluating the intervention’s effectiveness in more severe levels of depression could determine whether improving self-help via e-mail is only possible or effective for mild levels of depression. It is possible that the e-mails would not be sufficiently motivating for those who are more depressed and would not change self-help behaviour. It is also possible that improving self-help behaviour would have little or no benefit and that more powerful interventions are needed for more severe depression. Therefore, the current paper reports on a second trial of the Mood Memos intervention. In contrast to the original trial, participation by individuals with any level of depressive symptoms was allowed. We also admitted participants who were already receiving depression treatment, both for ethical reasons and because it is likely that the system would often be used this way in practice. The intervention could be well suited as an adjunct to cognitive behaviour therapy, as it could reinforce lessons learned during therapy. We expected that the active Mood Memo e-mails would reduce depressive symptoms more than the control e-mails overall, as well as in sub-samples of participants with case level major depression and with sub-threshold depression. We also hypothesised that participants allocated to the active Mood Memo e-mails would increase their usage of the promoted self-help strategies, relative to participants assigned to the control e-mails.http://clinicaltrials.gov/ct2/show/NCT01399502) and was approved by the University of Melbourne Human Research Ethics Committee  Participants were recruited to the study through the internet, as this allowed for broad participation, and was a successful recruitment technique in the first Mood Memos study d = 0.17) and the small effect size of unsupported internet-based treatment of depression (d = 0.25) The effect size of the Mood Memos intervention was expected to be small, given the small effect size found in the first Mood Memos study  and at the end of the intervention (6 weeks post-baseline).The primary outcome was depression symptom severity at post-intervention, assessed with the PHQ-9. The PHQ-9 assesses the frequency over the past two weeks of the nine Criterion A symptoms of DSM-IV Major Depressive Episode N = 1,602).Self-help strategy use was assessed pre- and post-intervention only. Participants rated how frequently over the past month they had used the 14 self-help strategies promoted in the Mood Memo e-mails to improve their depressive symptoms. Frequency of use was rated on a 5-category scale . Ratings of mean frequency of use of each strategy were calculated, with higher figures indicating greater use (range: 0 to 3). Use of each strategy was combined into a measure of total strategy use. This total strategy use scale ranged from 0 to 42 and had good internal consistency  at pre-intervention. In addition, at the mid- and post-assessment they were asked whether they had visited a health professional to help deal with depression during the previous 3 weeks. If yes, they were asked what treatments they had received, with multiple responses allowed: none, just a consultation; acceptance and commitment therapy; behaviour therapy; cognitive behaviour therapy; dialectical behaviour therapy; family therapy; mindfulness-based cognitive therapy; problem solving therapy; psychoanalytic psychotherapy; supportive counselling; psychotherapy but not sure which type; antidepressant medication; antipsychotic medication; anti-anxiety medication; mood stabilising medication; stimulant medication; medication but not sure which type. Examples of medications  were provided.Analyses were undertaken on an intention-to-treat basis. Any participants who were randomised but withdrew from the study were included in the analysis as randomised. Psychopathology and self-help use outcomes were evaluated using mixed models for repeated measures (MMRM) In the major depression sub-group, response to the intervention was compared between groups. The standard definition of clinical response on the PHQ-9 was used: a score of 10 or more at baseline, 50% improvement between pre- and post-intervention, and a post-intervention score of 9 or less Several potential predictors of attrition were explored. These were pre-intervention PHQ-9 score; baseline major depression; age; gender ; receiving treatment at baseline (yes/no); highest level of education (primary/secondary school or trade versus bachelor or postgraduate degree); history of depression (yes/no); history of bipolar disorder or psychotic disorder (yes/no); and adherence to the intervention . These predictors were entered into separate logistic regression analyses with presence of data at post-intervention as the outcome variable.d) were calculated by dividing the difference between the two group means at post-intervention by their pooled standard deviation. Effect sizes based on change scores were used where there were significant baseline differences. Effects were tested at the p<.05 level (two-tailed), except for multiple comparisons, where the p values were adjusted using Holm’s method Where means did not significantly differ between groups at baseline, between-group effect sizes (Cohen’s t (1719.3) = 2.78, p = .006, mean difference of 1.17 (95% CI: 0.34 to 2.00). Scores on the PHQ-9 at pre-intervention ranged from 0 to 27, but on average indicated moderately severe depression . The median number of symptoms experienced was 6 (range 0 to 9). Two-thirds of participants (66.0%) met criteria for a major depressive episode, this proportion was not significantly different between groups, χ2  = 2.81, p = .094. Mean scores on the K10 indicated a very high level of psychological distress There were 1,736 participants fully enrolled in the study. Participants ranged in age from 18 to 95 years, with a mean of 37.8 years. Most participants were female, less than a third of the sample possessed at least a bachelor degree, more than half reported a history of depression, and a third reported that they were currently receiving treatment for depression from a health professional. The two groups were well matched on sociodemographic characteristics at pre-intervention see . At pre-Participants were free to seek treatment for depression during the study. Of those who provided data at mid- or post-intervention, 43.6% (366/839) reported receiving treatment prior to the mid- or post-assessment. Overall, more than half  of these participants reported receiving treatment for depression at some point during the study (including pre-intervention). About a third of participants  reported receiving some form of medication, primarily antidepressants, during the study. A quarter of participants (215/839) reported receiving some form of psychotherapy during the study .2 (1) = 4.00, p = .045. There was large attrition from assessments at mid- and post-intervention (see 2 (1) = 1.50, p = .220. Reminders to complete assessments had a small effect on attrition. Of participants with post-intervention data, 64% completed assessments with no reminders, 21% completed after receiving one reminder, and 15% completed after receiving two reminders. Participants with data at post-intervention were significantly more likely to: have a history of depression OR = 1.73 (95% CI: 1.40 to 2.13) p<.001, have a history of bipolar disorder or psychotic disorder OR = 1.68 (95% CI: 1.05 to 2.70) p = .031, have a university education OR = 1.87 (95% CI: 1.51 to 2.33) p<.001, be older OR = 1.03 (95% CI: 1.02 to 1.03) p<.001, report being in treatment for depression at the beginning of the study OR = 1.96 (95% CI: 1.59 to 2.41) p<.001, and have read more than 4 of the intervention e-mails  p<.001; 9 to 12 e-mails, OR = 8.57 (95% CI: 6.41 to 11.5) p<.001). Although pre-intervention PHQ-9 score was not a significant predictor OR = 0.98 (95% CI: 0.97 to 1.00) p = .086, participants who met criteria for major depression were less likely to have data at post-intervention OR = 0.80 (95% CI: 0.65 to 0.98) p = .035. Gender was not a significant predictor, OR = 0.99 (95% CI: 0.77 to 1.27) p = .937.A minority of participants elected to stop receiving the intervention , with slightly more withdrawing from the control group than the active group (6.2% versus 4.1%), χtion see . The attF  = 3.14, p = .044 (see t (867.1) = 2.44, p = .015. Mean improvement in the active group was 0.91 points (95% CI: 0.18 to 1.65) more than the control group, d = 0.23 (95% CI: 0.09 to 0.37). The mean difference in improvement between pre- and post-intervention was not significant, t (692.4) = 0.75, p = .453, mean difference in improvement 0.35 points (95% CI: −0.57 to 1.28), d = 0.11 (95% CI: −0.06 to 0.27). A similar pattern of results was found for the K10. The interaction between group and time on the K10 was significant, F  = 3.94, p = .020, but differences between groups in change from pre-intervention to post-intervention were not significant, t (663.9) = 0.84, p = .399. However, the active group improved significantly more than the control group between pre- and mid-intervention, t (817.1) = 2.77, p = .006, mean difference in improvement 1.2 points (95% CI: 0.36 to 2.11), d = 0.22 (95% CI: 0.08 to 0.36). The interaction between group and time for the WSAS was not significant, F  = 2.84, p = .059, and there were also no significant differences between groups on the change from pre- to post-intervention, t (675.7) = 0.76, p = .445, d = 0.03 (95% CI: −0.13 to 0.20). Again, the active group showed more improvement than the control group from pre- to mid-intervention, t (833.7) = 2.33, p = .020, d = 0.12 (95% CI: −0.02 to 0.26). Data on adverse events were not collected in this study, however, a small proportion of participants (3.0%) recorded an individually reliable deterioration in PHQ-9 scores from pre- to post-intervention .The means and standard deviations of the primary and secondary outcomes at pre-, mid-, and post-intervention are shown in .044 see . HoweverF  = 1.42, p = .243 (see t (572.0) = 0.48, p = .634. The interaction between group and time on the K10 was not significant, F  = 2.71, p = .068, and differences between groups in change from pre-intervention to post-intervention were also not significant, t (572.0) = 0.89, p = .375. Lastly, the interaction between group and time for the WSAS was not significant, F  = 1.36, p = .258, and there were no significant differences between groups on the change from pre- to post-intervention, t (572.0) = 0.40, p = .692.Similar results were obtained when analyses were restricted to participants that provided complete data pre- and post-intervention . The MMRM interaction of group and time narrowly escaped significance, F  = 2.89, p = .057, the mean difference in improvement between pre- and post-intervention was not significant, t (410.4) = 0.95, p = .345, but the contrast between pre- and mid-intervention was significant, t (527.4) = 2.39, p = .017. The mean difference in improvement from pre- to mid-intervention was 1.19 points (95% CI: 0.21 to 2.17), d = 0.16 (95% CI: −0.01 to 0.34), but this was much smaller and non-significant between pre- and post-intervention, 0.60 points (95% CI: −0.65 to 1.85), d = 0.06 (95% CI: −0.15 to 0.27). Overall, mean PHQ-9 scores post-intervention were much reduced but still within clinical range . A larger proportion of participants in the active group  achieved a clinical response than the control group , but the intervention response rate was not significantly different . The number needed to treat to achieve one clinical response to the intervention was 19.3 (95% CI: 6.8 to ∞ to NNTH 23.8).There was a similar pattern of results on the PHQ-9 when the analysis was restricted to the 1,145 participants who met diagnostic criteria for major depression at baseline  than in the active group , t (394) = 2.17, p = .031. The MMRM interaction of group and time was not significant, F  = 0.50, p = .609. Mean difference in improvement between groups was not significantly different between pre- and post-intervention, t (156.5) = 0.95, p = .345, 0.47 points (95% CI: −1.10 to 2.04), d = 0.08 (95% CI: −0.25 to 0.41); or between pre- and mid-intervention t (191.0) = 0.99, p = .326, 0.6 points (95% CI: −0.60 to 1.81), d = 0.17 (95% CI: −0.12 to 0.46).A sub-group analysis was also conducted on participants who met criteria for sub-threshold depression at baseline (n = 396), to explore whether the first Mood Memos study results were replicated. Mean PHQ-9 scores at baseline were slightly but significantly higher in the control group  and control group , t  = −1.35, p = .177. An MMRM indicated significant differences over time between groups, F  = 22.6, p<.001. The active group improved by a mean of 2.63 points more than the control group (95% CI: 1.54 to 3.72) on the strategy use scale, d = 0.40 (95% CI: 0.23 to 0.56). Results from completers only were very similar, with significant differences over time between groups, F  = 20.2, p<.001, and a mean difference in improvement of 2.62 points (95% CI: 1.48 to 3.77), d = 0.40 (95% CI: 0.23 to 0.56). In addition to evaluating the effect of the intervention on overall usage of the promoted self-help strategies, the effect on individual strategies was explored to examine whether the e-mails had more effect on some strategies than others. d = 0.27 to d = 0.47).At pre-intervention there was no significant difference on the strategy use scale between the active group (SD = 4.0). However, the distribution was U-shaped, so the mean does not accurately summarise the distribution. The number of e-mail views was split into a 3-level ordinal variable: viewers of few e-mails (1 to 4), viewers of a medium number of e-mails (5 to 8), and viewers of most e-mails (9 to 12). Approximately two-fifths of participants (37.6%) read four or fewer e-mails, one fifth (20.6%) read 5 to 8 e-mails, and two-fifths (41.8%) read 9 or more e-mails. The proportion of participants in these categories did not significantly differ between groups, χ2 (2) = 1.16, p = .559. To explore the effect of adherence on depression outcomes, a linear regression was conducted using e-mail views categorised into three levels. Incremental (linear) and non-linear effects of email view category were evaluated. Adjusting for pre-intervention depression score and group allocation, e-mail views was a significant predictor of post-intervention depression score, B = −0.71 (95% CI: −1.38 to −0.04), p = .038, but the percentage of variance accounted for was very small when it was added to the model (ΔR2 = .006). The addition of a quadratic e-mail views term was almost significant B = 0.43 (95% CI: −0.02 to 0.87), p = .062, ΔR2 = .005. Adherence was also examined to see whether it was associated with clinical response in participants with major depression at baseline. In the active group, intervention response rates were 25.0%, 35.3% and 36.4% for those who read 1 to 4 e-mails, 5 to 8 e-mails, and 9 to 12 e-mails, respectively. This suggests some benefit from reading more than 4 e-mails, but little additional benefit from reading 9 or more. Whereas in the control group, response rates were 25.0%, 36.1% and 21.1%, for those who read 1 to 4 e-mails, 5 to 8 e-mails, and 9 to 12 e-mails, respectively.Both groups had a similar pattern of e-mail views. There was a sharp drop in the number of views between the first and second e-mail  and then a low rate of attrition for the remaining e-mails, with the twelfth e-mail viewed by 49.3% of the active group, and 49.9% of the control group. However, participants showed a mixed pattern of e-mail views, with some e-mails missed but later ones viewed. The mean number of e-mails viewed in the entire sample was 6.8 , and perhaps the anticipated small effect of the intervention on depression was overwhelmed by the larger benefits of depression treatments. It is also possible that the increases in self-help behaviours were too small, and that larger changes could be achieved by modifying the intervention. Some potential modifications include tailoring on dose, use of strategies, and stage of change for managing depression A sub-group of participants were highly adherent to the intervention, reading at least 75% of the e-mails, but there was also a significant proportion who read four or fewer e-mails. Adherence in web-based depression self-help interventions is often measured in number of modules completed, and evidence suggests a positive relationship between adherence and outcomes The finding that e-mail messages that promoted self-help behaviours for depression led to increases in those behaviours is encouraging. These results support other research showing periodic prompts and reminders to engage in health behaviours can be effective in improving health outcomes, at least in the short term Assessment attrition was substantial and associated with clinical history, level of education, intervention adherence and case-level depression at baseline. The e-mail reminders reduced attrition somewhat, but the high rate of attrition was not unexpected given the lack of participant supervision and ease of ignoring e-mail requests to complete assessments. The data were analysed with mixed models for repeated measures, which retains all available data from participants rather than deleting cases with incomplete data or replacing missing data with the last valid observation. These analyses were consistent with the results of completers only. The study was also limited in its reliance on self-report measures to assess depression and self-help behaviour. A clinical interview may have identified individuals with depression caused by a medication or medical condition, which would not have been amenable to intervention with self-help. Clinician-rated scales are also considered the gold standard in depression outcome research. Recent research suggests however that both clinician-rated and self-rated instruments should be used, because they each provide unique information and their estimates of treatment efficacy differ significantly Overall, the effects on psychopathology must moderate the encouraging results found in the first study. This is disappointing because the intervention required minimal resources to operate and could have been widely disseminated if it proved effective. Other, admittedly more intensive, internet-based interventions for depression are available and have been shown to be effective in several meta-analyses Checklist S1CONSORT checklist.(DOC)Click here for additional data file.Protocol S1Trial protocol.(PDF)Click here for additional data file.Active emails S1Screenshots of the active group emails.(PDF)Click here for additional data file.Control emails S1Screenshots of the control group emails.(PDF)Click here for additional data file.