Adjuvant psychological therapy (APT) is a newly developed cognitive behavioural treatment which has been designed specifically to improve the quality of life of cancer patients by alleviating emotional distress and inducing a fighting spirit. We report a phase I/II study which evaluates APT in routine clinical practice. A consecutive series of 44 outpatients with various cancers referred for psychiatric consultation and receiving APT at the Royal Marsden Hospital was studied. Standardised self-report questionnaires were used to measure anxiety, depression and four principal categories of mental adjustment to cancer, namely, fighting spirit, helplessness, anxious preoccupation and fatalism. Statistical comparisons between pre-therapy scores and scores after an average of five APT sessions revealed significant improvement in anxiety, depression, fighting spirit, anxious preoccupation and helplessness. Fatalism scores showed the same trend, but the changes were smaller. Patients with advanced disease showed as much improvement as those with local or locoregional disease. Present results indicate improvement in both psychiatric symptoms and mental adjustment to cancer associated with APT. Whether this association is causal remains to be determined by randomised controlled trials. Such a trial is in progress.


To pursue these findings, we sought to elucidate the mechanisms underlying target gene selectivity for HIF-1α and HIF-2α. Using chromatin immunoprecipitation to probe binding to hypoxia response elements in vivo, and expression of chimaeric molecules bearing reciprocal domain exchanges between HIF-1α and HIF-2α molecules, we show that selective activation of HIF-α target gene expression is not dependent on selective DNA-binding at the target locus, but depends on non-equivalent C-terminal portions of these molecules. Our data indicate that post-DNA binding mechanisms that are dissimilar for HIF-1α and HIF-2α determine target gene selectivity in RCC cells.Inactivation of the von Hippel–Lindau tumour suppressor in renal cell carcinoma (RCC) leads to failure of proteolytic regulation of the α subunits of hypoxia-inducible factor (HIF), constitutive upregulation of the HIF complex, and overexpression of HIF target genes. However, recent studies have indicated that in this setting, upregulation of the closely related HIF- However, there is increasing evidence for the functional non-equivalence of HIF-1α and HIF-2α, and in cancer, several recent studies have indicated that, at least in certain settings, they have contrasting effects on tumour growth TTGATTGGTAACTTCG 3′- and 5′-ACG aga tct(BglII) ATA GTT CTA CCA GGC 3′. Primers for the latter were 5′-ACT aga tct(BglII) TCT TCA TTG TGC TCA TCC CC 3′ and 5′ CGc cat gg(NcoI)T CGC CCG CAG AAT CGA GGT CC-3′. PHD3.HRE-SV40-luc was created by cloning the PHD3 first intron fragment into the multiple cloning site of pGL3promoter . A mutant HRE version of this reporter was created using site-directed mutagenesis with: 5′-CCACCCTCACGCAGCGacatTAGCCCTGTCACTCAGCC-3′ and a complementary oligonucleotide 5′ GGC TGA GTG ACA GGG CTA atg tCG CTG CGT GAG GGT GG 3′ (mutated nucleotides in lower case). Replacement of the SV40 promoter in pGL3promoter with the cloned PHD3 promoter gave rise to PHD3.HRE-PHD3pro-luc. PHD3pro-luc-PHD3.HRE was created by placing the PHD3 first intron fragment into a BamHI site downstream of the poly(A) signal in pGL3promoter using the following primers: 5′-Ggg atc c(BamHI)TC GAG TTG ATT GGT AAC TTC G-3′ and 5′-Ggg atc c(BamHI)AT AGT TCT ACC AGG C-3′. pCMV-β-galactosidase  and a 1 kb fragment of the PHD3 promoter upstream to the translation start site  were amplified by PCR from human genomic DNA using primers containing the appropriate restriction sites . Primers for the former were 5′-ATggtacc or hypoxia (1% oxygen and 5% carbon dioxide for 16 h) in an Invivo2 hypoxic workstation .The VHL competent cell lines used were Caki-1 . Hep 3B  and MCF-7 . The VHL defective cell lines 786-O , A498  and RCC4 (a gift from CHMC Buys) were all derived from renal clear cell tumours. All cell lines were maintained in Dulbecco's modified Eagles Medium supplemented with 10% fetal calf serum, 2 mα and HIF-2α were used as described previously  using Oligofectamine reagent  on day 1, cotransfected with siRNA duplexes (20–100 nM), 800 ng luciferase reporter plasmid and 200 ng pCMV-β-galactosidase plasmid using Lipofectamine 2000 Reagent (Invitrogen) on day 2, and harvested on day 3. In overexpression experiments, cells were cotransfected with 500 ng HIF-α expression plasmid (pcDNA.Hs.HIF-1α or pcDNA.Hs.HIF-2α), 300 ng HRE-linked luciferase reporter plasmid  and 200 ng pCMV-β-galactosidase plasmid using Lipofectamine 2000 reagent on day 1, and harvested on day 2.Two sets of siRNA duplexes targeting HIF-1μl 1 × passive lysis buffer (Promega) and assayed for luciferase activity according to the manufacturer's instructions. β-Galactosidase assay was performed to normalise for transfection efficiency.Cells were collected in 100 α , HIF-2α , CA9 (M75) . Horseradish peroxidase-conjugated goat anti-mouse secondary antibody  and ECL Plus system  were used for detection.Whole cell extracts from six-well plates were prepared in urea-sodium dodecyl sulphate (SDS) buffer, normalised for total protein content, resolved and transferred as previously described  and anti-HIF-2α (PM9) antibodies were used in the IP. PM14 and PM9 were raised by immunising a rabbit with a fusion protein consisting of glutathione- S-transferase fused to amino acids 445–553 of mouse HIF-1α or amino acids 357–439 of mouse HIF-2α, respectively. Both antisera cross react with the relevant human HIF isoforms. Dilution buffer and rabbit immunoglobulin were used as negative controls. The resulting DNA was analysed by PCR with primers flanking the HREs of PHD3 intron 1 and CA9 promoter  using standard ChIP protocols . Hep3B cpromoter .α and HIF-2α.For ChIP–Western analysis, urea–SDS lysis buffer was added directly onto the beads used for IP of DNA–protein complexes after the washing steps and boiled at 95°C for 5 min. After centrifugation at 4000 r.p.m. for 5 min, the supernatant was separated by polyacrylamide gel electrophoresis, transferred and blotted with mouse monoclonal antibodies against HIF-1α , HIF-1α protein with enhanced electrophoretic mobility (A498) or no immunoreactive HIF-1α protein (786-0). The concordance of results produced with independent sets of siRNA duplexes indicates that they are due to specific effects on the targeted isoform. Thus, PHD3 expression conforms to the pattern that has previously been described for HIF target genes such as VEGF and GLUT1  or HIF-2α (PHD3). Further analysis of immunoprecipitates from 786-O cells revealed enrichment of both HREs by the antibody to HIF-2α, but as expected neither HRE was captured in immunoprecipitates using the antibody against HIF-1α  implicated in DNA binding into the HIF-2α molecule did not enable activation of the CA9 gene. In contrast, replacement of the entire C-terminal HIF-2α sequence (amino acids 95–870) with the equivalent HIF-1α sequence (amino acids 96–826) was sufficient to convey full activity on the CA9 target gene. Comparison of the effects of the chimaera containing HIF-1α amino acids 96–826 with those containing less extensive C-terminal sequences  implicated in DNA binding into the HIF-1α molecule did not enable activation of the PHD3 gene, whereas replacement of the entire C-terminal HIF-1α sequence with the equivalent HIF-2α sequence (amino acids 95–870) was sufficient to convey full activity on the PHD3 target gene. So far the relationship between HIF-1α and CA9 and that between HIF-2α and PHD3 appeared to be reciprocal. However, further analysis revealed that whereas extension of the N-terminal HIF-2α sequence to amino acids 1–542 did not restore PHD3 activation to the chimaera, the HIF-2α C-terminal sequences 543–870 conveyed full activity, indicating that these sequences were both necessary and sufficient for HIF-2α activation of PHD3  promoter appears exclusively responsive to HIF-1α.To analyse further sequences conveying HIF-1α on the endogenous PHD3 gene and effects on the juxtaposed enhancer/promoter sequence in PHD3.HRE-PHD3pro-luc. We considered whether altering spacing between the promoter and enhancer might affect HIF-α selectivity. The PHD3 enhancer was therefore moved to a more distant position 3′ to the luciferase reporter gene in PHD3pro-luc-PHD3.HRE. Analysis of activity in transiently transfected RCC4 cells again showed that activity was dominantly mediated by HIF-1α rather than HIF-2α , siRNA suppression of the inactive isoform did not clearly enhance transcription, suggesting that competition for binding is not important at endogenous levels of the proteins, at least under the circumstances examined.Our findings indicate that this target gene selectivity is achieved by post-DNA-binding mechanisms. Chromatin immunoprecipitation experiments demonstrated non-selective binding for HIF-1α target CA9 and the HIF-2α target PHD3. First, the results of the HIF-1α/HIF-2α domain exchange experiments were non-reciprocal, and implicated non-equivalent sequences in activation of CA9 and PHD3 expression. Second, experiments directed towards defining the cis-acting DNA sequences at the CA9 and PHD3 loci that might mediate HIF-α isoform-selective responses yielded quite different results. Thus, for CA9, these experiments suggested that sequences at the minimal promoter could convey selective responses to HIF-1α. siRNA-mediated suppression of HIF-α isoforms indicated that the minimal CA9 promoter, like the endogenous gene, was specifically responsive to endogenous HIF-1α and not HIF-2α. Although some breakdown in selectivity was observed under conditions of overexpression by transient transfection, in comparison with other HREs, the CA9 promoter again appeared specifically responsive to HIF-1α, suggesting that specificity determining sequences in HIF-1α operate through direct or indirect interactions with factors binding the CA9 promoter.Further analysis suggested that the post-DNA binding mechanisms activating target gene expression are different for the HIF-1α. Rather siRNA suppression experiments revealed that the sequence was dominantly responsive to HIF-1α. This was observed irrespective of whether the PHD3 HRE was linked to the PHD3 promoter or a heterologous promoter, for different promoter/HRE spacings, and in both transiently and stably transfected RCC4 cells. Interestingly, in contrast with the lack of effect of siRNA-mediated suppression of HIF-2α, overexpressed HIF-2α powerfully activated the PHD3 HRE. This unexpected finding has recently been noted by others who have proposed that HIF-2α transcriptional activity is downregulated by a titratable repressor that is overcome by overexpression of the molecule , NF-κB essential modulator (NEMO) has been identified as a HIF-2α binding partner that promotes transcriptional activity (α with its endogenous target genes in RCC will require further investigation. Our studies provide a new focus for such investigations and indicate that they should be directed at post-DNA-binding mechanisms regulating HIF-2α target gene expression.A number of other recent studies have addressed possible mechanisms underlying differential activity of HIF-1


Patients with Parkinson's disease (PD) sometimes develop impulsive compulsive behaviours (ICBs) due to their dopaminergic medication. We compared 26 impulsive and 27 non-impulsive patients with PD, both on and off medication, on a task that examined emotion bias in decision making. No group differences were detected, but patients on medication were less biased by emotions than patients off medication and the strongest effects were seen in patients with ICBs. PD patients with ICBs on medication also showed more learning from negative feedback and less from positive feedback, whereas off medication they showed the opposite effect. The basal ganglia, the prefrontal cortex, the amygdala and the fusiform gyrus are all believed to be involved in face processing In this study we tested PD patients on and off their usual dopaminergic medication to examine the effects of impulsivity and dopamine in decision making in a task relying on emotional cues. Participants were asked to learn by trial and error, which of two faces (a happy and an angry face) was associated with a small monetary reward and then to pick that face as often as possible. In theory the facial expression should not influence decision making, however studies in healthy volunteers showed that subjects would select the happy face more often than the angry one, especially when learning was difficult A functional imaging study has shown that emotion bias correlates with activation in the anterior cingulate, which is known to have the greatest dopamine innervation in the cortex Impulsive compulsive behaviours (ICBs) such as pathological gambling and compulsive sexual behaviour are triggered by dopaminergic medication. Although some risk factors for ICBs and behavioural changes on metric tasks have been described Thus we speculated that PD patients with ICBs (PD+ICB) would be less biased by emotions in their “on” versus “off” state and hypothesised that controls would learn better on this task than PD+ICB patients.To assess the effects of dopaminergic medication on sensitivity to social bias in PD patients with and without ICBs.Twenty five PD patients without ICBs (PD−ICB) and 26 PD+ICB patients were recruited from the National Hospital for Neurology and Neurosurgery, Queen Square, London. All patients fulfilled the Queen Square Brain Bank criteria for the diagnosis of PD PD patients were tested before and after their usual anti-Parkinson medication in a counterbalanced sequence to account for order effects. All patients who were tested in their “off medication state” did not take their usual anti-Parkinson medication, including both L-dopa and any dopamine agonists, for at least 12 hours and performed the task between 8.00am and 9.00am. They were then retested in their “on medication” state the following day, usually mid mornings. Those patients who were tested “on medication” first performed this task usually in mid-morning when their motor symptoms were well controlled. They were re-visited on the following day prior to their medication for the second test. Results were compared with 16 controls who were matched for age to the PD+ICB group and who were tested over two days but did not take any anti-Parkinson medication. At the end of the study all participants received a modest honorarium depending on their final score .We used a probabilistic face decision task, which has been described previously All participants provided written informed consent according to the declaration of Helsinki and the study was approved by the UCLH Trust ethics committee.All data analysis was carried out in Matlab. Details about the statistical analysis have been reported elsewhere Analysis of variance was carried out on emotion bias, overall learning, and the effects of positive vs. negative feedback. In all cases, these parameters were computed for each subject, and then an ANOVA was run with the corresponding behavioural measure as a dependent variable. Emotion bias and overall learning were characterized by compiling a 2×2 contingency table for each subject. Each cell in the contingency table contained counts of choice patterns of the model and the observer. Based on the counts in the table, emotion bias was computed as: = 5.3, p = 0.008). Post hoc analysis showed that the PD−ICB group was older than the PD+ICB group (p = 0.009). There was no difference between controls and PD or PD+ICB patients (p>0.08). PD+ICB patients had a significantly younger disease onset relative to PD−ICB patients . There was no difference in LEU dose, disease duration or years of education between both patient groups. Further there was no group difference in UPDRS part 3 motor scores prior to (‘off-state’) and similarly no group difference after medication (‘on-state’). Significantly more PD−ICB than PD+ICB patients were treated with a dopamine agonist, which is in line with accepted clinical guidelines of managing an ICB in PD , thus all patients chose the angry picture more often in their ‘on’ compared to their ‘off’ state. There was no difference between groups in emotion bias  and no interaction between medication and group . Further, there was no effect of fraction correct performance (reward learning), when included as a covariate, on emotion bias . Thus, participants who had a larger emotion bias did not necessarily learn less. When the PD groups were analysed individually to examine medication effects, there was a significant decrease in emotion bias in the PD group on vs. off medication  but not in the ICB group . In contrast to the emotion bias, there were no effects of medication  or group  on overall learning , off medication  and ICB group off medication  and controls . The PD+ICB on medication group just missed significance . For overall learning performance, the PD−ICB on  and control  groups were above chance, but all other groups were at chance (p>0.156).Thus, all patients, except the PD+ICB on group, chose the happy face significantly more often than the angry face, given equivalent reward feedback.We next compared the PD−ICB and PD+ICB groups, off and on medication, to controls, pair-wise. However, there were no significant group differences in emotion bias (p>0.071). Following this, we carried out planned comparisons on emotion bias separately in each group, to see if each group was significantly affected by the facial expression . A signi1, 24 = 0.23, p = 0.629) or medication by feedback type interactions . However, in the PD+ICB group we found that there was a significant medication by feedback type interaction  but no main effect of medication . Thus, consistent with our previous study In the final analysis we split overall learning into parameters which separately measured learning from positive and negative feedback in the PD groups off and on medication . Results‘anticipated’ the correct choice. This anticipation may be mediated via the ventral striatum and anticipation of a conflict is processed via the anterior cingulate All patients chose the happy face more than the angry face, when expressions were matched for reward history, in their ‘off’ versus ‘on’ state. Furthermore, when examined pair-wise there were no differences in emotion bias or learning between the patient groups and the control group. This might be because participants generally were not able to learn and integrate the reward information effectively, since their behaviour did not correlate well with the ideal observer, which is a purely statistical model of how one should learn. When each group was analysed individually, we found that all groups, other than the PD+ICB group ‘on medication’, had a significant emotion bias and preferred the happy to the angry face. Our finding is consistent with previous studies using the same task Thus, PD+ICB patients may learn poorly in some conditions because they might use irrational estimates of correct responses, rather than trying to learn from previous trials.We found that both patient groups selected angry faces less than controls, although none of the groups differed significantly from controls. Patients with schizophrenia also select angry faces less often than controls, given equivalent reward evidence The nonapeptide oxytocin can decrease emotion bias in this task, specifically by increasing choices of angry faces when the evidence supports angry faces In an fMRI study of this task, larger modulation of blood-oxygen-level dependent (BOLD) responses by reward prediction error in the anterior cingulate and temporal parietal junction, were correlated with smaller emotion bias effects. Further those subjects that learned best in this task had the smallest modulation of BOLD responses by reward prediction error In summary, dopaminergic medication decreased sensitivity to social bias in both groups of PD patients, in the absence of group effects and none of the groups differed from matched controls. Our results might imply that PD+ICB patients ‘on medication’ are less affected by emotional feedback, which could be partly responsible for their difficulties integrating into society and following social norms. This is of interest in relation to the finding of increased schizotypy in PD with ICBs


The correct title and legend of Figure 3 presently appears with Figure 4. The correct, complete Figure 3 legend and title are:Time to negative NS1 antigenaemia.Kaplan – Meier survival analysis of time to resolution of NS1 antigenaemia by treatment group (CQ or placebo) and population; A) Intention to treat population and B) Per Protocol population. Two patients NS1 negative at enrollment were later positive (both in the CQ arm) and for the purposes of analysis were considered positive at the time of enrolment.doi:10.1371/journal.pntd.0000785.g004


The correct title and legend of Figure 4 presently appears with Figure 2. The correct, complete Figure 4 legend and title are:Time to fever clearance.Kaplan – Meier survival analysis of time to fever clearance by treatment group (CQ or placebo) and population; A) Intention to treat population and B) Per Protocol population. Three patients afebrile at enrollment developed fever later (1 in the CQ arm and 2 in the Placebo arm) and for the purposes of analysis were considered positive at the time of enrolment.doi:10.1371/journal.pntd.0000785.g002


The correct title and legend of Figure 2 presently appears with Figure 3. The correct, complete Figure 2 legend and title are:Time to resolution of viraemia.Kaplan – Meier survival analysis of time to resolution of plasma viraemia by treatment group (CQ or placebo) and population; A) Intention to treat population and B) Per Protocol population.doi:10.1371/journal.pntd.0000785.g003


Escherichia coli and evaluated for its biological activity.Interleukin 7 (IL-7) is a ϒc family cytokine involved in the homeostatic proliferation and maintenance of immune cells. In the present study, we report the expression of bovine IL-7 (bIL-7) in E. coli (BL 21 DE3). The expressed protein was purified by nickel-nitrilotriacetatechromatography, and the reactivity of the protein was confirmed by western blotting using monoclonal antibodies raised against human IL-7. The biological activity of expressed bIL-7 was evaluated by analyzing its effect on the expression of anuclear factor for activated T-cells c1 (NFATc1), B-cell lymphoma 2 (Bcl2), suppressor of cytokine signaling 3 (SOCS3) molecules in bovine peripheral blood mononuclear cells (PBMCs) by quantitative polymerase chain reaction. Ability of the expressed protein was also analyzed by its effect on phosphorylating signal transducer and activator 3 (STAT3) molecule by immunostaining in human embryonic kidney cells 293 (HEK293) cells.The sequence coding for bIL-7 (mature protein) was amplified from primary bovine kidney cell culture and cloned into pET28-a vector and expressed in The bIL-7 was able to induce the expression of Bcl2 and NFATc1expression in bovine PBMCs by 7 and 5-folds, respectively, whereas a 2-fold decrease was observed in the case of SOCS3 expression. Immunostaining studies in HEK293 cells using antihuman phospho-STAT3 showed activation and nuclear translocation of STAT3 molecule on bIL-7 treatment.E. coli expressed bIL-7 protein is biologically active. Considering the role of IL-7 in T-cell homeostasis and memory cell generation, this molecule can be used for enhancing the vaccine response and that has to be proved by further experiments.bIL-7 gene was successfully amplified, cloned, and expressed in a prokaryotic expression system. The biological activity study showed that the  Interleukin 7 (IL-7) is a vital cytokine for the homeostasis and development of both T- and B-lymphocytes. It is produced by intestinal epithelial cells, keratinocytes, hepatic tissues, peripheral blood dendritic cells, endothelial cells, smooth muscles, and fibroblast cells in human, however in mouse, thymic stromal cells, fetal thymus cells, bone marrow, spleen, kidney, keratinocytes, and fetal intestine are identified as the sites of IL-7 production . IL-7 isL, and MCI-1, which are also anti-apoptotic in nature [IL-7 activation promotes survival and proliferation of lymphocytes . The surn nature ,7. Supprn nature ,9. The mn nature . Very ren nature .The crystal structure of IL-7 receptor and their binding properties with IL-7 were well-studied in the case of human IL-7 .in vitro.Since there is no report on the characterization of bovine IL-7 (bIL-7), the present study reports the cloning of full-length sequence of bIL-7 and expression of the bIL-7 mature (bIL-7 mat) protein in prokaryotic expression system and evaluation of its biological activity All the procedures were carried out with prior ethical permission of Institute Bio-Safety Committee (IBSC) and Institute Animal Ethics Committee (IAEC), Indian Veterinary Research Institute, Hebbal, Bangalore. (Ref.No. F.8-56-Vol.II/RCSS/2014-15/dtd 8-06-2015).Bovine kidney cells were collected in normal saline from the local slaughter house, and bovine primary kidney cell culture was prepared. Total RNA was isolated from the primary bovine kidney cell culture using TRIZOL reagent .cDNA was prepared by reverse transcriptase reaction using oligo dT primers. The cDNA was used to amplify the IL-7 full-length sequence using IL-7L (5’-ggcgggtaccatgttccatgtttcttttagg-3’) and IL-7R (5’-ggcggcggccgctcagtgttctttaatgccc-3’) primers which were designed using IL-7 sequence available in the database (Gene Bank Accession No: BC142466.1). The purified polymerase chain reaction (PCR) product was ligated into EcoRV digested pcDNA (+3.1) vector. The cloned insert was subjected for sequencing by an automated sequencer, and the sequence was confirmed by comparing with known related sequences present in the database by NCBI blast tool.E.coli. Overnight grown cultures of four randomly selected recombinant colonies were inoculated to 10 ml Luria-Bertani broth medium and grown to optical density600= 0.6-0.8 at 37°C. Protein expression was induced with 1 mM isopropyl thiogalactoside (IPTG) for 5 h at 30°C. The pellet was lysed and analyzed by 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) gel for protein expression under denaturing conditions. After confirming the protein expression, the fusion protein was purified by nickel-nitrilotriacetate (Ni-NTA) chromatography  using denaturing conditions for eluting the protein by elution buffer containing 250 mM imidazole. Protein was partially refolded by reducing the molarity of urea in the washing buffer and different fractions of elute were analyzed by SDS-PAGE. The purified samples were ran on 10% SDS-PAGE and transferred to nitocellulose membrane for western blotting using human Il-7 specific monoclonal antibodies .bIL-7 mat sequence was amplified from the full-length IL-7-pcDNA plasmid using IL-7Lmat (5’-gcggatccgattgtgatattagaggtaaagac-3’) and IL-7R (5’-ggcggcggccgctcagtgttctttaatgccc-3’) primers. The PCR product was cloned into pET28a vector at BamHI and NotI sites to generate pET28-bIL-7 mat and transformed into DH5α cells. The recombinant plasmid pET28-bIL-7mat confirmed by restriction digestion analysis and expressed in BL21c DE3 strain of 6 cells were dispersed in each well in a six-well plate. About 100 ng of the recombinant protein was used per well for the biological activity assay. PBMCs were incubated with mature IL-7 for 3 h, and total mRNA was collected by TRIzol reagent . cDNA was synthesized using oligodT primers. Real time primers were designed for Bcl2 , NFATc1 , and SOCS3  molecules using Primer blast software of NCBI. ABI prism 7300 (Applied Biosystems) was used to carry out qPCR with conditions consisting of an initial step at 50°C for 2 min and 95°C for 2 min followed by 40 cycles at 95°C for 15 s and at 60°C for 1 min. The qPCR results were analyzed by relative quantification method [The biological activity of purified mature IL-7 was studied by its effects on expression of Bcl2, nuclear factor for activated T-cells c1 (NFATc1), and suppressor of cytokine signaling 3 (SOCS3) molecules. Peripheral blood mononuclear cells (PBMCs) were isolated from cattle blood using histopaque 1077 . Approximately 2 × 10n method , where g5 human embryonic kidney cells 293 (HEK293) cell per well were seeded in six-well plate and allowed to grow for 24-36 h. Cells were incubated with 10 μg/ml of purified E.coli expressed IL-7 mature protein for 30 min. After incubation cells were washed twice with 1 × phosphate buffered saline (PBS) and fixed with 3.7% formaldehyde in 1 × PBS  for 15 min. The plate was again washed twice with 1 × PBS and treated with 0.5% triton x-100 (1 × PBS) for 15-20 min to permeabilize the cell and nuclear membrane. Cells were washed twice with 1 × tris-buffered saline (TBS)-Tween  and blocked with 3%BSA (in TBS-Tween) for 1h. Washing was repeated thrice with 1 × TBS-Tween and incubated with the anti human phospho-STAT3 antibody (1:1000 in TBS-Tween) for 1 h at 37°C. After washing secondary antibody conjugated with fluorescein isothiocyanate (1:2000 in 1 × TBS-Tween) was added to the plates and incubated for 1h at 37°C. The plates were washed with 1 ×TBS-Tween and observed under fluorescence microscope.Immunostaining analysis of STAT3 molecule activation was carried out in a six-well culture plate. Approximately 1 × 10E.coli, and protein expression was induced by 1mM IPTG. On SDS-PAGE analysis, an extra band of 20 kDa was observed in induced cultures when compared with uninduced cultures (In this study, the recombinant plasmid was transformed into BL21c DE3 strain of cultures . Since tcultures , a singlcultures .A thick cultures .E.coli expressed bIL-7 was evaluated by assessing itseffect on the expression of Bcl2, NFATc1, and SOCS3 molecules. In this study, the effect of bacterial expressed IL-7 on these molecules was analyzed by qPCR by keeping GAPDH gene as an internal control. The results showed 7.2-fold increase in the expression of Bcl2, 4.8-fold increase in the NFATc expression, and 2.3-fold decrease in the expression of SOCS3 compared to the control in bovine PBMC culture (Biological activity of  culture .E.coli expressed IL-7 on phosphorylating STAT3 molecule is analyzed by immunostaining. HEK293 cells were incubated with 10 μg/ml of recombinant protein for 30 min. Later cells were processed for immunostaining according to the above-described procedure. Cells incubated with recombinant bIL-7 showed prominent fluorescence in the nucleus (The effect of  nucleus  and c, w nucleus  and f.IL-7 is believed to be the most important cytokine involved in the homeostatic proliferation of T- and B-cells. IL-7 signals through a receptor complex of IL-7Rα and γ chains . IL-7 is+T-cells [Homeostatic proliferation and survival of immune cells are mainly attributed to the function of IL-7. The survival effect of IL-7 is mainly elicited by up-regulating anti-apoptotic proteins such asBcl2-family proteins by the JAK3/STAT5 pathway. Expression of Bcl2 decreases in effector cells and increases to the maximum in memory cells, when compared to naïve CD8+T-cells . The exp+T-cells ,16. Rece+T-cells . A paralSOCS molecules are the physiological suppressors of cytokine signaling. They function as internal regulators of cytokine signaling, and many of the SOCS molecules are activated as a response to cytokine signaling. Because of their cytokine regulatory function, SOCS molecules are exploited by many infectious agents to escape from host immune system. It has been shown that IL-7 therapy augments IL-6 production by inhibiting SOCS3 activation by regulating FoxO transcriptional activity. Our resE.coli expressed IL-7 on phosphorylating STAT3 molecule is analyzed by immunostaining. In the present experiment, HEK293 cells incubated with IL-7 showed fluorescence in the nucleus indicating the nuclear translocation of the phosphorylated STAT3 molecules,whereasthe nucleus of the control cells did not show any fluorescence and remained restricted to the cytoplasm. On activation, STAT molecules would be translocated into the nucleus and elicit effects at the transcription level. Phosphorylation of STAT molecule is one of the modes of action of IL-7. STAT5 is documented as the major candidate for the JAK/STAT signaling pathway by IL-7. In the development of B-cells, especially pre-B-cells depend on IL-7-mediated STAT3 signaling. Studies revealed that STAT3 deficient pre-B-cells are highly susceptible for apoptosis [27-thymocytesin adult mice and promote the capacity of γδ27- to produce IL-17, which help in the polarization of T-cells. This selectivity of IL-7 on γδ27-thymocytes to produce IL-7 mainly owes for its capacity to activate STAT3 in such cells [The effect of poptosis . IL-7 inch cells . Like STch cells .Imperative role by IL-7 in T-cell memory prompted the molecule to be tested for its adjuvanted effect in its DNA and protein forms. It showed increased T-cell response against hepatitis C virus infection  and aviaE.coli expressed bIL-7 matare biologically active. Uponstimulation with the recombinant bIL-7, there was an increase in the mRNA levels of NFATc1 and Bcl2 and a decrease in the expression of the SOCS3 molecule in PBMCs. Use of IL-7 protein and IL-7 encoding DNA constructs are reported to be useful against cancer and many viral infections. Biologically active bIL-7 expressed in E.coli will be a good candidate to evaluate further it as a therapeutic agent as well as an adjuvant to the vaccine in bovines.The present study shows that JL- The research work mentioned in this article is a part of the Ph.D research work of the first author and first author carried out all the work mentioned in this article. NV - The immunostaining of HEK cells to analyze the biological activity of the bIL-7 protein. JHD - The real time PCR assay described in the present study. GRR- The entire work mentioned in this article was carried out under the guidance of the fourth author.


Diffraction gratings are an essential optical component of high-power, short-pulse lasers. The maximum output of high-power pulsed lasers is always determined by laser resistance of gratings and this resistance is strongly dependent on the local near electric field intensity in the grating structure. We presented a novel method of reducing electric-field-enhancement in metal-dielectric grating by designing asymmetric grating ridge while maintaining high diffraction performance. Compared with the common isosceles trapezoidal grating, the grating with asymmetric ridge got a 0.04% reduction of diffraction efficiency in TE polarization at 1053 nm incident wavelength but a 21.3% reduction of maximal electric-field-enhancement in grating structure. This method can be applied to any surface-relief gratings to reduce the electric-field-enhancement for improving the laser induced damage threshold (LIDT) of grating and supporting the grating-based chirped pulse amplification (CPA) system to develop into higher peak-power levels. Metal gratings were firstly and commonly used as PCGs in CPA systems because metals can naturally reflect light with broadband high efficiency3, but they were limited to the laser induced damage threshold (LIDT)2. Compared with metal gratings, all-dielectric gratings feature negligible losses and much higher LIDTs5 but cannot fulfill the broader bandwidth specifications6 and the mechanical stress will increases significantly when the number of layers and size become large7. For avoiding these disadvantages mentioned above, mixed metal-dielectric grating with high diffraction efficiency (DE) and broad bandwidth is a viable candidate for the realization of PCG with high LIDT8.The pulse compression grating (PCG) is one of the most critical components of a high-power grating-based chirped pulse amplification (CPA) laser system9 and recently we found that this grating was initially damaged at the grating ridge by femtosecond laser in transverse-electric (TE) polarization. The initial damage was occurred at the location where the enhancement of the electric field intensity  is highest in the grating as shown in Fig. et al.5. Steve Hocquet et al. also reported this phenomenon in multilayer dielectric (MLD) grating and they improved the LIDT from 2.9 J/cm2 up to 4.8 J/cm2 in normal beam by illuminating the MLD grating from in TE to transverse-magnetic (TM) polarization with a reduction of electric-field- enhancement from 1.38 to 1.16. But they stressed that MLD grating have been designed so far to be used in TE polarization because the DE is much lower in TM polarization10. So polarization conversion seems not a practical method and direct reducing electric-field-enhancement in grating is required to improve the LIDT of the grating used with femtosecond laser. Bonod and Neauport reported a drop of the maximum field intensity by increasing the groove width11, but Liu et al. reported a similar method but decreasing the groove width12, so the method of adjusting the groove width seems to be uncommon because it strongly relies on the specific grating itself. In any case, achieving high LIDT while maintaining high diffraction performance is a significant and challenging work for supporting the grating-based CPA system to develop into much higher peak-power levels. In this paper, we present a metal-dielectric grating solution of reducing electric-field-enhancement in the structure and achieving high DE at the same time.A broadband metal-dielectric grating with a sandwich-ridge was designed and fabricated in our previous work9. For the sake of simplicity, the grating ridge with single material was set to be rectangle in the beginning. So our grating contains a rectangle grating ridge (height hg), a residual layer (thickness tr), a matching layer (thickness tm), a high reflective Ag layer (thickness tAg) and the substrate. The material of the grating ridge and the residual layer is Ta2O5 because high-refractive-index material can lower the etching depth and increase the DE and also can be compared to the material in sandwich grating structure mentioned in Fig. 2 is set to match the phase for achieving higher DE. We focus on designing the grating with broadband high DE around 1053 nm wavelength. Multi-parameter optimization is carried out in our design because multiple parameters of the grating influence the DE and bandwidth of grating. These parameters included the period of the metal-dielectric grating Λ, the duty cycle f, the height of grating ridge hg and all the thicknesses . The incident angle was fixed at -1st reflective Littrow-mount constraint by g = 0.439 μm, tr = 0.163 μm, tm = 0.025 μm and tAg = 0.2 μm. The calculated -1st reflective DE spectrum in TE polarization and the schematic structure of the grating are shown in Fig. 13. So we sliced the rectangle ridge into 10 thin planar slabs and these slabs were randomly shifted with a slight displacement away from the center. The calculated DE spectrum and the schematic structure of the deformed grating are shown in Fig. The asymmetric distribution of electric field intensity see Fig.  is resul15. So we re-optimized the metal-dielectric grating into isosceles trapezoid shape to go a step further for describing our designing method. Based on our previous experiments15, the base angle of β in the grating was mostly measured near 80° with a ±10° range, so the isosceles base angle of β was set to 80° for convenience. The period of grating Λ was fixed at 0.667 μm as the same as the rectangle one for keeping consistent working situation. The trapezoidal grating ridge was sliced into 40 slabs to increase the accuracy of simulation. The re-optimized parameters of the isosceles trapezoidal metal-dielectric grating were: f = 0.72, hg = 0.59 μm, tr = 0.236 μm, tm = 0.178 μm and tAg = 0.2 μm. The calculated DE of the -1st order reflected spectrum in TE polarization and the schematic structure of this grating are shown in Fig. Design of isosceles trapezoidal grating ridge with base angle of β rather than the rectangular shape can lower the difficulty of fabrication based on our previous worksL and βR) were added to describe our grating shape and they were set to be 70° and 90° respectively when the incident light irradiated from left top. The two angles 70° and 90° are the results from the optimization process, which are separated ±10° from 80° and this equal difference will cause the ridge distortion in a certain direction. Compared with the isosceles trapezoidal grating ridge with 80° base angle, the non-isosceles grating can be considered that the sliced slabs were integrally shifted a small displacement opposite to the incoming wave. The other parameters of the asymmetric grating were optimized with slight changes: f = 0.7, hg = 0.6 μm, tr = 0.23 μm. The calculated -1st reflective DE spectrum in TE polarization and the schematic structure of this asymmetry grating are shown in Fig. et al., they got a 39.58% increasement of LIDT by a 18.96% diminution of electric-field-enhancement10, so we have confidence to believe that the LIDT of the metal-dielectric grating can be improved by designing asymmetric grating ridge because of these obvious reductions of electric-field-enhancement in grating.We want to design specific asymmetric grating shape to modulate the distribution of electric field intensity of the grating while maintaining high DE. According to the result from Fig. 16 to find minimum of merit function and the merit function was taken as a mean-error expression around the considered wavelength range N is the number of wavelength discrete points and 17 was employed to calculate the DE and the distribution of the enhancement of the electric field intensity. The grating bandwidth can be optimized depended on the wavelength range in the merit function. Using evolutionary algorithms such as GA has been proved to be a straightforward and efficient method to optimized grating based on our previous works18 and GA is also widely used in fiber optics19, three-dimensional printings20 and other optical components designing21.The optimization is obtained by the genetic algorithm (GA)9. In coating process, a chromium film was deposited on the substrate first to enhance the adhesion of metal film and substrate, and all layers were deposited by electron beam evaporation. The coating sample was annealed at 350 °C in air to release internal stress for avoiding delamination in the cleaning process. In lithography process, the grating ridge pattern of photoresist was generated from the interference of two monochromatic collimated beams of ultraviolet (UV) light. In etching process, the grating structures were transferred from the photoresist to the dielectric layers by reaction ion beam etching technology and cleaned by hydrochloric/peroxide mixture combined with mega-sonic cleaning method. Besides, the asymmetric grating ridge can be fabricated by specific slant-etching technology, which is common in slanted grating fabrication based on the studies by Tapani Levola et al.22 and Kalle Ventola et al.23. In the damage test procedure, each test site was only exposed to a single shot, and different sites were irradiated with the different laser fluence. The fluence was gradually decreased until the damage didn’t happened. The irradiated sites were preliminarily judged to be damaged or not with the aid of charge-coupled device (CCD) and the details of damaged spots were carried out by SEM.The metal-dielectric grating mentioned in Fig. All data generated or analysed during this study are included in this published article.26. This design method can be applied to any surface-relief gratings to reduce the electric-field-enhancement for improving the LIDT of grating and supporting the grating-based CPA system to develop into much higher power levels.In this paper, we presented a novel method of reducing electric-field-enhancement in metal-dielectric grating by designing asymmetric grating ridge. We firstly demonstrated that restricted distortion of grating ridge will not influence much on the DE, which means that grating can be designed with unusual shape while maintaining high DE. Associated with the initial damage at the grating ridge where the enhancement of the electric field intensity is highest in the structure, we modified our optimized isosceles trapezoidal metal-dielectric grating into non-isosceles shape with two base angles for modulating the distribution of electric field intensity in the grating. Compared with the isosceles trapezoidal grating, the grating with asymmetric ridge got a 0.04% reduction of DE in TE polarization at 1053 nm central incident wavelength but a 21.3% reduction of electric-field-enhancement in grating structure. This result indicated that it’s valid to design specific asymmetric grating shape to modulate the distribution of electric field intensity of the grating while maintaining high DE. Asymmetric gratings can be not only used to reduce laser damage but also used for other applications such as near-to-eye display technology and waveguide technology


A strong and anisotropic interfacial coupling between the charge transfer pairs is demonstrated. The van der Waals heterostructures exhibit pressure dependent sensitivity with a high piezoresistance coefficient of −4.4 × 10−6 Pa−1, and conductance and capacitance tunable by external stimuli (ferroelectric field and magnetic field). Density functional theory calculations confirm charge transfer between the n-orbitals of the S atoms in BEDT–TTF of the BEDT–TTF/C60 layer and the π* orbitals of C atoms in C60 of the P3DDT/C60 layer contribute to the inter-complex CT. The two-dimensional molecular van der Waals heterostructures with tunable optical–electronic–magnetic coupling properties are promising for flexible electronic applications.Two-dimensional van der Waals heterostructures are of considerable interest for the next generation nanoelectronics because of their unique interlayer coupling and optoelectronic properties. Here, we report a modified Langmuir–Blodgett method to organize two-dimensional molecular charge transfer crystals into arbitrarily and vertically stacked heterostructures, consisting of bis(ethylenedithio)tetrathiafulvalene (BEDT–TTF)/C Two-dimensional van der Waals heterostructures are of interest due to their unique interlayer coupling and optoelectronic properties. Here authors develop a Langmuir-Blodgett method to organize charge transfer molecular heterostructures with externally tunable conductance and capacitance and broadband photoresponse. Currently, 2D heterostructures are predominantly based on inorganic complexes, especially chalcogenides with strong vertical chemical bonds1. In this context, the urgent demand of flexible nanoelectronics and optoelectronics calls for a novel generation of organic heterostructures held by vdW forces for both vertical and horizontal orientation7. Among the organic complexes, the superior optoelectronic properties and the lack of interlayer screening effect for the 2D donor and acceptor CT molecular crystals, as well as the charge density wave induced long-range vdW force in the polarized structure, is a promising candidate to enable 2D CT molecular heterostructures with tunable optical–electronic–magnetic coupling behavior at the atomic level17.Two-dimensional (2D) heterostructures based on weak interlayer van der Waals (vdW) interaction with the lack of superficial dangling bonds afford multiple degrees of freedom for the creation of new high-quality 2D heterojunctions and superlattices without the constrains of lattice parameters, enabling customized and tunable optical–electronic–magnetic properties2. Therefore, it is critical to investigate the coupling behavior between two pairs of different CT layers, which holds promise to show unique coupling behavior distinct from traditional inorganic counterparts due to the inter-conversion between singlet and triplet CT states under external stimuli combined with the internal hyperfine interaction and spin-orbit coupling of organic semiconductors. Moreover, the combination of the distinct properties from different layers is not only complementary but also will enhance the performance of the whole system18.The atomically regulated interface of 2D heterostructures affords an ideal platform to understand the heterointerfacial coupling behavior. However, the conventional 2D heterostructures are composed of two layers of opposite carrier type with CT interaction1. Moreover, the choice of lattice-matched substrate is needed for the growth of 2D materials, which limits the scalable preparation20. Thus, a universal method for the preparation of organic 2D vdWHs is in urgent demand. On the other hand, the understanding and control of heterointerfacial coupling behavior for the organic vdWHs is of key importance towards the further design and applications of the nano-devices. Among all the approaches to control the interfacial state, extrinsic charge injection by ferroelectric field generated from flexible P(VDF-TrFE) polymer has drawn much attention. The large polarization of the crystallized ferroelectric polymer, P(VDF-TrFE) induces a net polarization of organic CT components with enhanced optoelectronic and even magnetoelectric performance24.To prepare 2D vdW heterostructures (vdWH), conventional chemical vapor deposition approach is widely used. However, the weak interlayer interaction between the monolayer 2D film and the substrate leads to the island growth rather than continuous monolayers60) acceptor layer (DTC60) and bis(ethylenedithio)tetrathiafulvalene (BEDT–TTF) donor with C60 acceptor layer (ETC60). The interfacial coupling can be tuned in over a large range by external fields (ferroelectric and magnetic) with strong enhancement of current and capacitance, due to the coupling across two CT pairs along the vertical and horizontal orientations.Here, we report the large scale assembly of 2D CT heterostructures with controlled orientation and unique physicochemical properties, consisting of vertically stacked poly (P3DDT) donor with fullerene  solvent with large surface tension, good spreading ability, and the low vapor pressure of 1,2-dichlorobenzene (DCB) solvent for ETC60 and DTC60 are all crucial for the assembly and stacking of 2D heterostructures  vibration of C60 in the ETC60-DTC60 heterostructure nanosheets compared to that in single ETC60 and DTC60 nanosheets confirms the existence of coupling between the ETC60 and DTC60 nanosheets , forming the segregated stacking 2D ETC60 nanosheets with the distance between each layer close to 6.46 Å28. The P3DDT layers self-organize through the π-π interactions16. The distinct structure of P3DDT and C60 molecules lead to the segregated stacking of P3DDT and C60 in the 2D DTC60 nanosheets, where the closest distance between the backbone of P3DDT and C60 molecules is 8.08 Å16. Each ETC60 nanosheet crystalline unit contains six molecules of BEDT-TTF and four molecules of C60, while that of DTC60 contains two molecules of P3DDT and two molecules of C60. The orthorhombic crystalline unit cell of ETC60/DTC60 has lattice parameters of a = 26.71 Å, α = 90°, b = 32.51 Å, β = 90°, c = 58.90 Å, γ = 90°. The horizontal stacking of the molecular chains on water surface forms a stable structure, which can also be verified by the following anisotropic properties. The CT interaction mainly exists along the vertical orientation, while π–π stacking and interchain interaction dominate the horizontal orientation.The crystallized 2D nature of such heterostructures is further confirmed by transmission electron microscopy and selected-area electron diffraction patterns Fig. . The shi60-DTC60 heterostructure devices. The optical, band alignment and anisotropic photoresponse properties are shown in Fig. 60 can be attributed to −5.68 and −4.37 eV, respectively. In addition, the HOMO and LUMO levels of the DTC60 nanosheet are −6.23 and −4.46 eV, respectively. The energy offset between ETC60 and DTC60 nanosheets facilitates the CT interaction for the coupling between ETC60 and DTC60 nanosheets , where h is planck’s constant, c demonstrates the speed of light, R is photoresponsivity, e represents the charge of an electron, λ refers to the wavelength of the irradiated light. The EQE for the horizontal and vertical orientations are 41% and 155%, respectively. According to the photocurrent formulaθ is the fitted slope related to the trapping and recombination process for the photogenerated charge carriers32. The θ values for the horizontal and vertical orientations are 0.961 and 0.959 under light intensity lower than 7 mW/cm2, which is close to one unit, revealing that monomolecular recombination is the dominant recombination process at low light intensity , we performed first principles density functional theory (DFT) calculations. The electronic density of states (DOS) and the partial density of states (PDOS) of the ETC60-DTC60 are shown in Fig. 60-DTC60 heterostructure results in shifting of the LUMO level closer to the Fermi energy. This decreases the HOMO-LUMO gap, and thus, contributes to the CT in the system. The HOMO is comprised mainly of the 2p states of S atoms in BEDT-TTF molecules of ETC60, while the 2p states of C atoms in C60 molecules of DTC60 dominate the LUMO band  in the heterostructure. There is also a significant contribution of the intra-complex charge transfer within the ETC60, but with a higher HOMO-LUMO gap than the inter-complex CT state  and I(0) is the current with and without magnetic field. The MC along vertical orientation is larger than that along horizontal orientation, and increases with the increase of magnetic field under dark  is much longer than that of singlet exciton (~ ps)38, triplet excitons contribute to the formation of dipoles, and thus the capacitance can be changed by magnetic field, i.e., magneto-capacitance effect  and C(0) are the capacitance under light illumination and dark, respectively , the heterostructure can also demonstrate magneto- and photoexcited capacitance. However, as discussed in the inset of Fig. 60 nanosheets and ETC60 nanosheets is much weaker than that of the opposite stacking. Consequently, C% for magneto-capacitance and photoexcitation is lower than 1% and 4%, respectively ark Fig. . It alsoark Fig. . Magnetiect Fig. . Consequect Fig. . The ampely Fig. . Photoex60 and DTC60 nanosheets, confirming the tuning ability of the CT states by ferroelectric field. Two different device structures are compared to reveal the mechanism of interface coupling. For the first structure, the stacking sequence from bottom to top is P(VDF-TrFE) layer, ETC60 nanosheet and DTC60 nanosheet, respectively  layer and DTC60 nanosheet, respectively  layer on the bottom of ETC60 nanosheet due to the screening of CT interaction between ETC60 and DTC60 nanosheets by the inserted P(VDF-TrFE) layer  ferroelectric field influences the behavior of dipoles, leading to the change of dielectric behavior. As shown in Fig. 60 and DTC60 nanosheets, it also shows the same enhancement behavior  ferroelectric polymer with high electrostatic field are applied to tune the interfacial coupling. The ferroelectric field from the P(VDF-TrFE) layer enhances the CT density and MC effect Fig. . The MC ely Fig. . The seqyer Fig. , reveali40. Two approaches are proposed to demonstrate the flexible electronic behavior of such 2D CT vdWHs, including tension strain and compress strain along the horizontal orientation. The inset of Fig. , ε is strain. The maximal gauge factor is 8.5. The flexibility can also be expressed by the piezoresistance coefficient41 according toX is the stress, −6 Pa−1 below a strain of 4.3%, and it decreases to −4.1 × 10−6 Pa−1 under higher strain. The piezoresistance coefficient of such 2D heterostructures is much larger than that of one dimensional nanowires42. The repeatability of the mechanical flexibility is confirmed by the pressure dependent electronic behavior  (P3DDT) donor with a fullerene acceptor layer and bis(ethylenedithio)tetrathiafulvalene donor with a fullerene acceptor layer into scalable free-standing sequential stacking vdWHs. The coupling between two CT layers endows the heterostructure with broadband fast photoresponse of milliseconds. The anisotropic packing structure leads to a much larger coupling extent of charge transfer along the vertical orientation than that of horizontal orientation, leading to anisotropic optical–electronic–magnetic stimulated optoelectronic properties. Moreover, the ferroelectric field effect from P(VDF-TrFE) is applied to tune the coupling and charge transfer in the heterostructures. The DFT calculations confirm the charge transfer between the n-orbitals of the S atoms in BEDT-TTF of ETC60 (Sigma-Aldrich Co.) was added to above solution and stirred at room temperature for 2 h. After centrifuged at 6000 rpm for 20 min, the supernatant solution was isolated and used for the preparation of the film. For P3DDT/C60 solution preparation, 5 mg/mL P3DDT (Ossila Ltd.) was dissolved in 1,2-DCB at 70 °C for 3 h. Then, 5 mg/mL C60 was added into the solution and stirred at room temperature overnight.5 mg/mL BEDT-TTF  was dissolved in 1,2-dichlorobenzene  solvent, and stirred at 80 °C for 3 h. Then, 5 mg/mL C60 solution was dropped slightly onto the surface of 20 mL distilled water/DMF mixed solution (volume ratio of 1:1) in a glass vessel and dispersed homogeneously. After several hours of slowly evaporation of 1,2-DCB solvent, thin two-dimensional nanosheet forms. The preparation method for P3DDT/C60 nanosheet is the same, except that only 20 μL solution is needed. The preparation details for the heterostructure devices, the preparation of P(VDF-TrFE) layer and the flexible devices on PDMS substrates for mechanical characterization are discussed in Supplementary information.In total 50 μL BEDT-TTF/CTopography images were taken by Bruker dimension icon atomic force microscopy (AFM) under tapping mode with Si probe at a scanning rate of 1μm/s. The radius and resonant frequency are 25 nm and 13 kHz, respectively.CHI 422 Series Electrochemical Workstation was used to get current and voltage signals. Driel’s 50–150 W Research Arc Lamp Sources was used to output simulated solar light. In addition 365 nm, 650 nm, and 850 nm light comes from UVL-21 compact UV lamp, THORLABS red light and THORLABS LIU 850 A near-infrared light, respectively. The signal was obtained at a sample interval of 0.001 s.60 and DTC60 solution were self-assembled onto the working electrode and dried at 40 °C in glove box to form uniformly coated nanosheets. The measurement step was 25 mV/s. The HOMO level was obtained from the equation: EHOMO = [−exp(Eonset (vs.Ag/AgCl)−Eonset (Fc/Fc + vs.Ag/AgCl))]−4.8 eV. The LUMO level was calculated from the obtained bandgap from the optical spectra.Cyclic voltammetry experiment was carried out in a three-electrode cell with glassy carbon electrode as the working electrode, Pt electrode as counter electrode, and Ag/AgCl electrode in saturated KCl water solution as the reference electrode. All the electrodes were cleaned and blown dry by nitrogen gas before use. The 0.1 M tetrabutylammonium tetrafluoroborate (Sigma Aldrich) was used as the electrolyte and dissolved in dehydrated acetonitrile (ACN). The stability of Ag/AgCl electrode was checked by ferrocene (Sigma Aldrich) as internal standard substance. The ETCCapacitance signal from 40 Hz to 10 M Hz was captured by Agilent 4294a Precision impedance equipment at 0.2 V and room temperature. Samples were collected by 16047E fixture. P(VDF-TrFE) ferroelectric layer was polarized by Keithley 2400 source meter at 15 V for 10 s.−1. Pressure was loaded by finger print. The pressure through finger was weighed and calibrated by a balance.Tension stress was loaded by Instron 5944 Single Column Tabletop Low-Force Universal Testing System at a speed of 0.5 μm s43. We use a generalized gradient approximation (GGA) with the Perdew–Burke–Ernzerhof (PBE)44 parameterization of exchange correlation energy functional with ultrasoft pseudopotentials45. An energy cutoff of 30 Ry was used for truncating the plane wave basis set to represent wave functions. Our recent work on ETC60 crystals gives excellent result with these parameters46. The structures were relaxed until the magnitude of the Hellman–Feynman force on each ion became smaller than 0.03 eV/Å. Brillouin zone integrations were carried out with a uniform small mesh of k-points. The Grimme parameterization47 was used to include the vdW interactions. For the purpose of this study, the heterostructure comprising of BEDT-TTF/C60 (ETC60) and P3DDT/C60 (DTC60) complexes (monolayer structure) was built using the Avogadro48 and Gauss View5 packages49. For the ETC60 section of the heterostructure, the crystal coordinates were used from the Cambridge Crystallographic Data Centre50. The monolayer structures of the ETC60 and DTC60 complexes were formed by removing the structures of DTC60 and ETC60, respectively from those of the heterostructure. The heterostructure contains total of 690 atoms out of which 396 belongs to ETC60 and remaining 294 associated with DTC60, respectively.Our calculations are based on first-principles density functional theory (DFT) as implemented in the Quantum ESPRESSO codehttps://doi.org/10.6084/m9.figshare.5005331.v1 Remaining data that support the findings are available from the corresponding author upon reasonable request.The data that support the findings of this study are available from the online data repository Figshare with identifier Peer Review FileSupplementary Information


We report rabies virus transmission among solid organ transplantation recipients in Changsha, China, in 2016. Two recipients were confirmed to have rabies and died. Our findings suggest that more attention should be paid to the possibility of rabies virus transmission through organ transplantation for clinical and public health reasons. In 2016, Zhou et al. reported a case of rabies virus transmission in China that was probably a result of organ transplantation , where he exhibited anemophobia, convulsions, limb rigidity, and hypersalivation. The patient was moved to hospital B (days 7–14) in Changsha. At admission, some examination findings indicated a possibility of viral encephalitis  Table 1.On day 14, the patient was transferred to hospital C, where he became comatose and was declared brain dead. Permission was granted for organ donation, because no specific pathogen had been detected and China’s organ transplant policy allows for organ donations in cases of viral encephalitides without laboratory-confirmed pathogens. Rabies virus–specific binding antibodies were also not detected in the serum samples. The donor had possibly been exposed to rabies but had not been vaccinated against rabies . The patient’s kidneys and liver were removed for transplantation in hospital D in Changsha on the following day.Kidney and liver transplantations were performed on December 10, 2015. Two female recipients in hospital D received the kidneys, and an 8-month-old girl in hospital E in Shanghai, China, received the liver. No other organs were collected or transplanted from this donor. The surgeries were all uneventful, and the recipients were discharged after transplantation. However, all 3 recipients were eventually readmitted to the hospital with complex symptoms . On the 40th day after transplantation, the left kidney recipient (29 years of age) revisited hospital D, reporting worsening right lower quadrant abdominal pain and vomiting. In the next 3 days, she successively had anemophobia, shortness of breath, chest tightness, anxiety, insomnia, and hypersalivation. During hospitalization, her blood pressure was as high as 248/148 mm Hg. On her 7th day in the hospital, she became comatose and then died. On the 43rd day after transplantation, the right kidney recipient (age 47 years) developed aches in her lower limbs, insomnia, and a decreased appetite. She was readmitted to hospital D the next day. On the 3rd day after admission, she exhibited hydrophobia, anemophobia, photesthesia, and fatigue. She showed agitation, dyspnea, and hypersalivation on the 4th day; she became comatose and died 1 day later. The liver recipient was readmitted with pneumonia on the 34th day after transplantation and died of asphyxia and multiple organ failure within 5 days. This patient did not show any signs or symptoms of rabies or encephalitis. None of the recipients had been exposed to potentially rabid animals or had been vaccinated previously for rabies online Technical Appendix). Both kidney recipients tested positive for rabies virus (. Both kiIn the past 10 years, rabies transmission by solid organ transplantation has been described occasionally worldwide (Epidemiologic information about donor and the 2 kidney transplant recipients in a rabies outbreak associated with solid organ transplantation, Changsha, China, 2015–2016.


Kidney transplantation is now a viable alternative to dialysis in HIV-positive patients who achieve good immunovirological control with the currently available antiretroviral therapy regimens. This systematic review and meta-analysis investigate the published evidence of outcome and risk of kidney transplantation in HIV-positive patients following the PRISMA guidelines.Searches of PubMed, the Cochrane Library and EMBASE identified 27 cohort studies and 1670 case series evaluating the survival of HIV-positive kidney transplant patients published between July 2003 and May 2018. The regimens for induction, maintenance therapy and highly active antiretroviral therapy, acute rejection, patient and graft survival, CD4 count and infectious complications were recorded. We evaluated the patient survival and graft survival at 1 and 3 years respectively, acute rejection rate and also other infectious complications by using a random-effects analysis.At 1 year, patient survival was 0.97 , graft survival was 0.91 , acute rejection was 0.33 , and infectious complications was 0.41 , and at 3 years, patient survival was 0.94  and graft survival was 0.81 .With careful selection and evaluation, kidney transplantation can be performed with good outcomes in HIV-positive patients. Traditionally, human immunodeficiency virus (HIV)-positive patients (HIV+) has not been considered to be good candidates for solid-organ transplantation for the poor prognosis of HIV patients. However, with the introduction of antiretroviral combination therapy (cART), the survival of HIV+ patients have been great improved. While the frequency of Acquired Immune Deficiency Syndrome (AIDS)-related events has consequently decreased, mortality due to organ failure has become a significant concern.The initial attempts at kidney transplantation (KT) in HIV+ patients led to poor outcomes, but better results occurred with the availability of highly active antiretroviral therapy (HAART) , 2.In this scenario, KT started to be proposed as a treatment even as “standard-of-care” for end-stage renal disease (ESRD) in selected HIV+ patients .A multicentre study in the USA found that the survival rates for HIV+ recipients fall between those reported for older KT recipients and for all recipients in the American national database .Despite these encouraging results, many issues still need to be addressed. Among the more relevant are the elevated incidence of acute rejection (AR), lower patient survival (PS) and graft survival (GS), and the hurdles caused by the interaction of immunosuppressive and antiretroviral (ARV) drugs. We conducted a systemic review and meta-analysis to determine the effectiveness of KT in the presence of HIV. Specifically, we examined PS and GS, AR and infectious complications in HIV+ patients who have undergone KT.The study design of a systematic review and meta-analysis was chosen to define the published evidence of the effectiveness of KT in HIV+ patients. The study followed the Preferred Reporting Items for Systematic Reviews and Meta-Analysis (PRISMA) statement standards . Our revWe searched the Medline (1966 to June 2018), EMBASE (1974 to June 2018), and Cochrane Controlled Trials Register databases to identify studies that referred to KT in HIV+ patients; we also searched the reference lists of the retrieved studies. The following search terms were used: KT, HIV+, AIDS. A combination of subject headings and keywords for KT, HAART, HIV+ recipient, allograft survival, antiretroviral therapy, donor selection, ESRD and immunosuppression was used for the literature search.Cohort studies and case–control studies were all eligible for inclusion if they reported outcomes of KT in HIV+ patients. Studies reporting outcomes shorter than 12 months post transplantation and transplantation occurring before HAART were introduced were excluded. Articles were independently assessed by 2 reviewers (X Z and WR X) according to the predetermined eligibility criteria. Any disagreement between reviewers was resolved by discussion with a third reviewer (XP H).All data were extracted independently by 2 reviewers (X Z and WR X) onto a Microsoft Excel spreadsheet , and any discrepancies were resolved by consensus. The following information was collected for each study: the study country, sample size, inclusion criteria, exclusion criteria, induction and maintenance immunosuppression, HAART regimen, mean CD4 T-cell counts (CD4 counts) pre-transplant and post-transplant, infectious complications, post-transplant neoplasia, PS and GS at 1 and 3 years, and AR rate. In order to analyse data of Infectious complications (IC), all infections requiring hospitalization were registered.The quality of each study used for the meta-analysis was assessed based on the Newcastle–Ottawa-Scale (NOS) for cohort studies . The evaWe undertook the descriptive analyses to identify the number of studies with relevant data, the countries where the studies were conducted, and other population attributes. The transplant outcome data were pooled using different transformations according to their different normal distribution conditions.The data for PS, GS, AR, IC at 1 year and PS at 3 years were analysed using log transformation.The data for GS at 3 years were analysed using arcsine transformation.The transformed data were combined to estimate the pooled percentages with 95% confidence intervals using a random-effects model.2 [http://www.R-project.org; version 2.9.0), using the software libraries ‘meta’ and ‘metafor’, for the meta-analysis and meta-regression models, respectively.The transformed data were combined to estimate the pooled percentages with 95% confidence intervals using a random effects model  and preson by I2 . StatistThe search strategy identified 86 citations Fig. , and amoDetailed characteristics of all included studies are provided in Table Regarding immunosuppression and rejection, most of the patients received antibody induction therapy with different regimens containing basiliximab, daclizumab, antithymocyte globulin (ATG) or methylprednisolone. The maintenance regimens were mainly composed of cyclosporin A (CSA), mycophenolate mofetil (MMF), tacrolimus (TAC) and steroids, which were the same as the maintenance regimens for HIV-negative patients.Mean CD4 counts were steady in most of the patients. As we observed, in all the cohorts with available data (22/27 cohorts), mean CD4 counts pre-TX and post-TX were greater than 200 cells/μL, and even elevated post transplantation.Pneumocystis jirovecii for at least 6 months.Prophylaxis against opportunistic infection was common in most studies as follows: patients received ganciclovir or valganciclovir for cytomegalovirus and trimethoprim/sulfamethoxazole, dapsone or atovaquone for Kaposi’s sarcoma and skin cancer were the most observed post-transplant neoplasia. As we showed in Table Each of the 27 studies included in the meta-analysis was assessed by the NOS to investigate the risk of bias within the studies. Table 2 = 36%) of patients survived, while 94%  of patients survived at 3 years.Twenty-seven studies reporting PS at 1-year post KT included PS estimate to post KT for 1429 patients; however, only nine studies including 509 patients reported PS at 3 years. The results of the analysis are shown in Figs. 2 = 69%) of grafts had survived, and GS subsequently declined to 0.81  at 3 years.Twenty-six studies including 1391 patients reported GS at 1-year post KT, and nine studies including 509 patients reported GS at 3 years. The results of the analysis are shown in Figs. 2 = 60%) of patients had AR.Twenty-five studies including 1051 patients reported AR post KT at 1 year, and the results of the analysis are shown in Fig. 2 = 59%) of patients had IC.Nineteen studies including 584 patients reported IC post KT at 1 year; the results of the analysis are shown in Fig. To our knowledge, this is the first systematic review and meta-analysis of such a large scale to report the outcomes of KT in HIV+ patients. We review and meta-analysis the outcomes in HIV+ KT patients, and looks at the 1- and 3-year GS/PS and AR rate.The availability of cART has made KT a feasible treatment for selected HIV+ patients with ESRD, with outcomes somewhat inferior to those observed among the overall population of KT recipients , 9–11.KT is now a viable treatment for select patients with HIV and ESRD. Moreover, the high incidence of morbidity and mortality resulting from cardiovascular issues in HIV+ patients , 13, as However, data regarding long-term outcomes and comparisons with appropriately matched HIV− patients are still lacking.Locke et al. analysed 510 adult KT recipients with HIV matched 1:10 with HIV− controls. They found that HIV− and HIV mono-infected KT recipients had similar GS and PS, whereas HIV/HCV co-infected recipients had worse outcomes .Izzo et al. found that the survival rate of patients was 82.1% and functioning grafts was 71.4% , and a rStock et al.  reportedIn our analyses, at 1 year, PS was 0.97 , GS was 0.91 , and at 3 years, PS was 0.94 , GS was 0.81 .One of the most challenging goals in solid-organ transplantation is to tailor the immunosuppressive regimen for each individual patient to minimize immunosuppression while still preventing AR. Opportunistic infections and malignancies often attributed to immunosuppression itself remain a significant cause of death after transplantation. In the field of HIV+ organ transplantation, finding a balanced approach to immunosuppression is even more critical.Currently, the vast majority of KT patients receive induction immunosuppression, which has been shown to greatly reduce the risk of rejection and improve PS and GS . As showGuidelines from the Kidney Disease: Improving Global Outcomes transplant working group recommended anti-IL2R as the first-line treatment for patients at low risk for rejection and ATG for those at high risk .Despite being the standard of care for most HIV patients, the use of induction immunosuppression for HIV+ patients, particularly ATG, remains controversial.On the one hand, HIV+ patients have high rates of rejection and thus stand to benefit significantly from induction. On the other hand, the risks posed from prolonged lymphocyte depletion are of major concern given that HIV+ patients are perceived to already threaten T cell populations and reduced immunity, both states that are associated with an increased risk of opportunistic infections.A recent study showed that ATG induction was associated with long-term impairment of T cell function and related infections, even after the patients had normalized CD4 counts . This fiThe incidence and severity of IC following transplantation are largely dictated by the recipient’s capacity for immune reconstitution. A study by Suarez et al. indicate that ATG-induced CD4 lymphopenia can be prolonged, and even at 1 year post transplant, a substantial proportion of patients has CD4 counts < 200/μL . The basA study by Bossini et al. showed that in HIV+ KT recipients treated with basiliximab and maintained on a calcineurin inhibitor (CNI)-mycophenolic acid (MPA)-based regimen, early corticosteroid withdrawal was associated with a very high incidence of AR and that kidney function was worse in patients with rejection .However, in contrast to the studies above, in a large national cohort of 830 HIV+ KT recipients, Kucirka found wide variation in the use of induction immunosuppression, with > 30% of HIV+ patients receiving no induction compared with only 20% of their HIV counterparts. Therefore, the study indicated that the use of induction, including the lymphocyte-depleting agent ATG, was not associated with an increased risk of infections. Despite the fact that induction recipients were at higher risk for AR, the researchers observed lower rates of delayed graft function (DGF), AR, graft loss and days spent hospitalized in the first year after KT as well as a trend towards lower mortality. They suggested that the benefits of induction immunosuppression to prevent graft rejection in HIV+ KT recipients far outweigh the perceived risk of increased infections. Because the study had the largest sample size to date and the cohort was nationally representative rather than a select study population, this study claims to be more credible. Furthermore, the authors accounted for confounding and treatment selection bias, which previous studies did not do, and they did this using inverse probability of treatment weighting (IPTW), a method that allowed them to adjust for many clinical and demographic factors even when modelling relatively rare outcomes such as graft loss and death .A high risk of AR is a well-known concern in HIV-infected kidney graft recipients. With regard to rejection, most studies observed a higher number of events in HIV+ patients than in HIV− patients. AR may occur as a result of immune dysregulation and the continuous inflammatory state of HIV+ recipients, in whom immunogenicity is increased following allograft implantation , 29.Vicari et al.  evaluateIn the study, the incidence of treated AR was higher in the HIV+ group, and the incidence of biopsy-confirmed AR was numerically higher in this group. Additionally, even though an identical incidence of antibody-mediated AR occurred, the incidence of steroid-resistant rejection was numerically higher in the HIV group. Many reports have revealed an elevated incidence of AR in HIV+ recipients, varying between 31% and 55% , 9, 10, However, Malat et al.  describeGathogo et al.  reportedIn addition, the elevated incidence of acute cellular rejection has been recently hypothesized to partially occur a result of an infiltration of inflammatory cells that occurs in response to tubular cell infection by HIV .A study by Malat showed a relatively higher incidence of mixed rejection in HIV+ recipients compared with that reported for non-HIV transplant recipients. A donor terminal serum creatinine greater than 2.5 mg/dL predicted mixed rejection and was associated with poor outcomes. Donor selection and optimization of immunosuppression may be critical in these patients . Even if2 = 59%), and the rate of incidence of IC observed in this study in HIV+ KT patients is in line with the frequencies reported in a study by Stock et al. [During the first decades of the renal transplantation era, a serious IC developed in up to 70% of patients following transplantation, resulting in fatal outcomes in as many as 11% to 40% of cases . In a reHowever, the long-term patient and graft outcome of the whole cohort were not influenced by HIV status but were adversely influenced by infections, as survival was diminished in patients having at least one infection.Furthermore, one-third of HIV+ KT recipients in a study by Ailioaie et al. did not have any episodes of infection, and repeated infections were not frequent. More importantly, the rate of incident infections was not different between the HIV+ and HIV− matched groups.As experience with transplantation in HIV+ patients grow, significant drug–drug interactions between ART and maintenance immunosuppression have been identified as a major clinical challenge.Post-transplant management of HIV infection with protease inhibitor (PI) and nonnucleoside reverse transcriptase inhibitor (NNRTI)-based ART is complicated by reciprocal drug interactions with immunosuppressive therapy, especially CNI, because of inhibition or induction of P450 cytochrome enzymes. Co-administration of PIs with CNIs requires significantly decreased CNI doses and prolonged dosing intervals to avoid supratherapeutic trough levels.Despite appropriate CNI dose adjustments, variations of drug serum levels are difficult to control and have been linked to increased graft rejection in HIV+ KT recipients , 39.In a study of 150 HIV+ KT patients, the largest to date, higher-than-expected rates of rejection were reported  [In a study by Rosa et al., patients receiving PI-containing regimens had lower PS at 1 and 3 years than patients receiving PI-sparing regimens—85% vs. 100% (p = 0.06) and 82% vs. 100% (p = 0.03), respectively .The increased risk of AR in HIV+ individuals has been largely attributed to reduced exposure to immunosuppressive agents due to drug–drug interactions with ART , 41, 42.This observation might be due to the effects of PI on tacrolimus levels, considering that the overwhelming majority of these patients were on a PI-containing regimen and that more than half had tacrolimus levels above target at the time of infection. PI could also influence the net state of immunosuppression by increasing the level or effect of other immunosuppressants, such as prednisone and mycophenolate.The most important finding in the present study is the association between PI use and adverse outcomes, namely, reduced 3-year PS and GS, and increased risk of serious non-opportunistic infections. These observations remained true in analyses restricted to patients receiving nucleoside reverse transcriptase inhibitor (NRTI) “backbone”; thus, even after excluding the potential influence of other agents included in the ART regimen, PI continued to be associated with poor outcomes.However, the use of NNRTI or tenofovir disoproxil fumarate (TDF) did not influence GS. Tenofovir alafenamide (TAF) is a new formulation of tenofovir associated with less kidney (and bone) toxicity . WhetherIn a large single-centre study of HIV+ KT recipients conducted by Boyle et al. , treatmeGiven that specific recipient characteristics, such as hepatitis B co-infection and certain HIV mutations, continue to make TDF-based regimens the most likely to provide adequate viral suppression post-transplant, despite observational data for nephrotoxicity in the non-transplant population.However, given the limitations of this study, TDF should be reserved for patients who have limited ART options and should be used very cautiously in the KT population, with appropriate dose adjustment and surveillance of kidney function, including kidney biopsy when indicated. Substituting TAF for TDF in KT patients is reasonable, but it should be noted that no data are yet available on long-term kidney outcomes with TAF in KT and non-KT recipients in the setting of both preserved and reduced glomerular filtration rate (GFR).Since their introduction in 2007, integrase strand transfer inhibitors (INSTIs) have been proposed as preferred post-transplant ART because of a favourable pharmacologic profile with decreased potential for drug interactions , 42, 46.Recently, Alfano et al., reported that preferred drug included raltegravir and dolutegravir for INSTI class, maraviroc for CCR5 receptor antagonist, lamivudine for NRTI, and rilpivirine for NNRTI, which offered advantage of having no drug interactions .In summary, we believe that INSTI or CCR5-based therapy should be the preferred ART in patients with HIV who undergo KT, primarily because of decreased drug–drug interactions with immunosuppressive medications such as CNIs, enabling easier monitoring of immunosuppressive medications and superior graft outcomes. However, larger and more controlled trials are needed to better assess the long-term outcomes of INSTI-based therapy to elucidate factors related to GS other than direct reciprocal drug interactions.In conclusion, this systematic review and meta-analysis demonstrated that with careful selection of patients and multidisciplinary evaluation, KT can be performed with good outcome in HIV+ patients. Moreover, with the advent of INSTI-based cART regimens, drug–drug interactions between cART and immunosuppressants have been dramatically reduced. Nevertheless, further studies are needed to optimize immunosuppressive therapy regimens for HIV+ patients, with the aim of reducing the high rate of AR after transplantation. Furthermore, this review still has its limitations, such as lack of sufficient studies, possibility of some overlapping patient cohorts, short of comparator. And we are also looking forward to other novel papers as more and more studies regarding KT of HIV+ patients.


The beneficial effects of silicon and its role for plants are well established; however, the advantages of silicon nanoparticles over its bulk material are an area that is less explored. Silicon nanoparticles have distinctive physiological characteristics that allow them to enter plants and influence plant metabolic activities. The mesoporous nature of silicon nanoparticles also makes them good candidates as suitable nanocarriers for different molecules that may help in agriculture. Several studies have shown the importance of silicon nanoparticles in agriculture, but an overview of the related aspects was missing. Therefore, this review brings together the literature on silicon nanoparticles and discusses the impact of silicon nanoparticles on several aspects of agricultural sciences. The review also discusses the future application of silicon nanoparticles in plant growth, plant development, and improvement of plant productivity. Equisetaceae family cannot survive in nutrient solutions lacking silicon  were successfully employed as an optical sensor for melamine detection. Similarly, Liu et al. (In recent decades, environmental sensing technologies based on metallic nanoparticles have gained enormous attention due to their extraordinary sensing resolution and sensitivity to analytes under optical detection. Developing a nanoparticle-based sensing device has been proposed due to the surface area of nanoparticles and the availability of surface anchored bioactive molecules for analyte detection (Hache et al. u et al.  preparedu et al. . TherefoNanotechnology is a promising area of interdisciplinary research that opens avenues in several fields, such as medicine, pharmaceuticals, electronics, and agriculture. This article presents the potential of Si-NPs in agriculture and brings together the literature relevant to the use of nanoparticles as pesticides, fertilizers, herbicides, genetic and drug transfer agents, soil improving agents, and sensors for soil analysis. Studies show that Si-NPs have the potential to revolutionize the existing technology used in various sectors, such as agriculture and plant biotechnology. Silicon nanoparticle-mediated targeting of biomolecules would be useful for developing new cultivars that are resistant to various biotic and abiotic factors. These nanoparticles can provide green and eco-friendly alternatives to various chemical fertilizers without harming nature. Thus, Si-NPs may have concrete solutions to many agricultural problems regarding weeds, pathogenicity, drought, crop yield, and productivity.


Juvenile dermatomyositis (JDM) is the most common inflammatory myopathy in childhood and a major cause of morbidity among children with pediatric rheumatic diseases. The management of JDM is very heterogeneous. The JDM working group of the Society for Pediatric Rheumatology (GKJR) aims to define consensus- and practice-based strategies in order to harmonize diagnosis, treatment and monitoring of JDM.The JDM working group was established in 2015 consisting of 23 pediatric rheumatologists, pediatric neurologists and dermatologists with expertise in the management of JDM. Current practice patterns of management in JDM had previously been identified via an online survey among pediatric rheumatologists and neurologists. Using a consensus process consisting of online surveys and a face-to-face consensus conference statements were defined regarding the diagnosis, treatment and monitoring of JDM. During the conference consensus was achieved via nominal group technique. Voting took place using an electronic audience response system, and at least 80% consensus was required for individual statements.Overall 10 individual statements were developed, finally reaching a consensus of 92 to 100% regarding (1) establishing a diagnosis, (2) case definitions for the application of the strategies (moderate and severe JDM), (3) initial diagnostic testing, (4) monitoring and documentation, (5) treatment targets within the context of a treat-to-target strategy, (6) supportive therapies, (7) explicit definition of a treat-to-target strategy, (8) various glucocorticoid regimens, including intermittent intravenous methylprednisolone pulse and high-dose oral glucocorticoid therapies with tapering, (9) initial glucocorticoid-sparing therapy and (10) management of refractory disease.Using a consensus process among JDM experts, statements regarding the management of JDM were defined. These statements and the strategies aid in the management of patients with moderate and severe JDM. Juvenile dermatomyositis (JDM) is the most common inflammatory myopathy of childhood and a major cause of morbidity and mortality among patients with pediatric rheumatic diseases . The manThe PRO-KIND  initiative is a sub-committee of the Society for Pediatric Rheumatology (GKJR) in Germany and Austria and aims to define consensus-based plans to harmonize diagnostic and treatment approaches. This initiative was started since it was perceived that children with juvenile rheumatic diseases in Germany and Austria often are either treated too late or not treated with the most up-to-date therapeutic regimens. To meet this challenge, the goal of the PRO-KIND initiative is to foster the use of harmonized diagnostic and treatment plans with defined targets , 26. PreAs a next step, the goal of the PRO-KIND working group on JDM was to harmonize previously identified patterns into plans and statements.www.surveymonkey.com). Nine pediatric rheumatologists participated in an initial face-to-face meeting in January 2016. There was consensus that the expert panel should also include expert pediatric neurologists. Therefore, 2 pediatric neurologists with special expertise in managing JDM (W. M-F. and U.S.) were also invited to join the group, forming an extended interdisciplinary expert panel.The JDM working group was formed in April 2015, initially comprised of 12 pediatric rheumatologists with clinical expertise in the management of patients with JDM. Identification of project goals took place via a formal online survey using the web-based program SurveyMonkey  and a non-voting dermatologist experienced in the care of children with JDM participated in a final face-to-face consensus meeting on January 13, 2017. The extended expert panel overlapped with but was not identical with the members of the initial working group. Prior to the consensus conference, the voting members had received access to various essential data via sciebo, a cloud service hosted by the state of Northrhine-Westphalia. The data provided included detailed results of previous online surveys, previous meeting protocols, essential literature obtained via an extensive literature search , including the 112 papers that were scored by SHARE . The proThe expert status of the panel members is demonstrated by the fact that centers represented in the panel contributed more than half of all patients  with JDM documented within the National Pediatric Rheumatic Disease Database (“Kerndokumentation rheumakranker Kinder und Jugendlicher”).Regarding the diagnosis of JDM, there was consensus that the conventional Bohan and Peter criteria should be modified. In particular, there was consensus that typical magnetic resonance imaging (MRI) findings represented an important finding for establishing a diagnosis of JDM. Furthermore, electromyography was removed from the list of findings. The finding of symmetric proximal muscle weakness was modified to also include myalgia as a possible criterion Table . ConcernIt was felt to be important to define a minimum dataset that should be routinely collected on all patients with at least moderate JDM Table . The groThe group defined a treat-to-target strategy, which means treatment to be adjusted according to the achievement of previously defined targets Table . It was There was consensus that all patients with JDM should receive supportive therapy including physical therapy to prevent contractures, muscle weakness, de-conditioning and disability. Early return to sports activities is encouraged as long as it is safely possible. Effective sun and/or ultraviolet light protection using avoidance, textile protection or sunscreen is strongly suggested. Additional supportive therapies may include hydroxychloroquin (HCQ), vitamin D or calcium supplementation Table . It was Different glucocorticoid regimens were preferred by experts within the panel. Based on the results of a previous survey , 3 glucoNo consensus was found concerning a specific tapering protocol for glucocorticoids. However, the group defined landmarks for the glucocorticoid taper, i.e. the aim to discontinue glucocorticoids after 12 months of therapy, and, in case of initial high-dose oral glucocorticoid therapy, PDN doses of 50, 25 and 12.5% 2 months, 4 months and 6 months after initiation of therapy, respectively  therapy, such as AZA, CSA or MMF. Furthermore, there was consensus that high-dose IVIG may be used in conjunction with DMARD therapy, especially in case of severe JDM Table . HoweverThe expert group considered refractory disease to be present if the predefined treatment targets were not reached. Furthermore, patients with longstanding (i.e. not new-onset) JDM may be treated according to strategies outlined here. The group considered two options to be viable under these circumstances: (1) Additional therapy with another DMARD, specifically, AZA, calcineurin inhibitos (CSA or tacrolimus), CYC, IVIG, MMF, RTX or TNFi, or (2) changing therapy to one of these medications. However, there was no consensus on which specific sequence of medications would be preferred. In order to reflect this uncertainty, the medications or medication classes are simply listed alphabetically Table .The statements developed are condensed in Fig. Based on data on the current clinical practice in Germany we defined consensus-based statements and strategies supporting the diagnosis and the management of JDM. The expert panel consisted both of pediatric neurologist and rheumatologists with all of them having practical experience in managing JDM. The group supports the use of “adapted” Bohan and Peter criteria for the diagnosis of JDM, including the use of specific MRI findings as a Bohan and Peter “equivalent” criterion, similar to what others have proposed in the past . There aSince there is a relative lack of data from randomized controlled clinical trials (RCTs), and since it is unlikely that large-scale RCTs for JDM will take place, one possible solution to gather more information on the efficacy or inefficacy of various treatments may be to collect “real life” data on disease outcome, i.e. to pursue comparative effectiveness research . An impoThese harmonized consensus strategies overall fit rather well into the above-mentioned framework set up by the CARRA consensus treatment plans and the SHARE recommendations which had been developed previously or simultaneously, respectively. However, there are important differences in clinical practice in Germany when compared to this framework. For example, practitioners in Germany have a strong preference to choose a glucocorticoid regimen consisting of ongoing intermittent IVMP therapy in combination with low-to-moderate dose glucocorticoids instead of conventional daily high-dose glucocorticoid regimens . There iThere are several limitations to the PRO-KIND consensus strategies for JDM. First, the concepts outlined here may not represent the optimal way to manage JDM but rather represent current clinical practice among JDM experts in Germany and are based on a low level of evidence. Second, the statements and resulting strategies do not represent treatment protocols or “To Do”-lists. Management of patients with JDM will remain highly variable. Third, the field is constantly evolving: Some MSAs that were only recently reported in children, for example, anti-3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMGCR) antibodies, are not included in our list but may be useful to determine in some patients, especially in case of refractory myositis , 46–48. In summary, we have developed consensus-based strategies for the diagnosis, monitoring and treatment of JDM by the harmonization of current clinical practice with the overarching goal is to improve the outcome of patients with JDM. Concerning therapy, we are placing an emphasis on a treat-to-target strategy using internationally accepted improvement criteria rather than on individual medications, fitting well into existing frameworks. As others have done previously, we define a minimal dataset to regularly collect in patients with JDM in order to allow comparative effectiveness research. We conclude that, due to the rarity of the condition, international collaboration will be critical in order to improve the outcome of patients with JDM. We believe that this work will add to the armamentarium of providers managing patients with JDM worldwide.