Rhabdomyolysis is an uncommon side effect of trabectedin which is used for the second line therapy of metastatic sarcoma after anthracycline and ifosfamide failure. This side effect may be due to pharmacokinetic interactions caused by shared mechanisms of metabolism involving the cytochrome P450 (CYP) system in the liver. Here, for the first time in literature, we describe the unexpected onset of heavy toxicity, including rhabdomyolysis, after the fourth course of trabectedin in a patient with retroperitoneal liposarcoma who at the same time was taking an alternative herbal medicine suspected of triggering this adverse event.This is the case of a 56 year old Caucasian man affected by a relapsed de-differentiated liposarcoma who, after the fourth cycle of second-line chemotherapy with trabectedin, complained of sudden weakness, difficulty walking and diffuse muscle pain necessitating complete bed rest. Upon admission to our ward the patient showed grade (G) 4 pancytopenia and a marked increase in liver lytic enzymes, serum levels of myoglobin, creatine phosphokinase (CPK) and lactate dehydrogenase. No cardiac or kidney function injuries were present. Based on these clinical and laboratory features, our conclusive diagnosis was of rhabdomyolysis induced by trabectedin.Aronia melanocarpa) daily during the last course of trabectedin and in the 2 subsequent weeks.The patient did not report any trauma or muscular overexertion and no co-morbidities were present. He had not received any drugs during treatment with trabectedin, but upon further questioning the patient informed us he had been taking a folk medicine preparation of chokeberry . Thus our case underlines the importance of understanding rare treatment-related toxicities such as trabectedin-induced rhabdomyolysis and the possible role of the drug-drug interactions in the pathogenesis of this rare side effect. Furthermore, this report draws attention to a potential problem of particular concern, that of nutritional supplements and complementary and alternative drug interactions. These are not widely recognized and can cause treatment failure. Ecteinascidia turbinate. Trabectedin was approved in Europe in 2007 as a single agent for second-line therapy of soft tissue sarcomas, and its activity is mainly relevant in liposarcoma and leiomyosarcoma [Despite aggressive surgery and adjuvant radiotherapy, more than half of patients with soft tissue sarcoma develop a locally advanced or metastatic disease . UnfortuWhile adverse reactions to trabectedin treatment such as neutropenia and liver function test alterations are well-known, more education and data are needed on some of the rare side effects mediated by this drug, such as the potential induction of rhabdomyolysis. A recent comprehensive safety analysis confirmed the uncommon occurrence of this side effect and also pointed out the need to avoid pharmacokinetic interactions with concomitant drugs caused by shared mechanisms of metabolism by the cytochrome P450 (CYP) system in the liver .We describe here the unexpected onset of heavy toxicity including rhabdomyolysis after the fourth course of trabectedin in a patient with retroperitoneal liposarcoma who at the same time was taking an alternative herbal medicine suspected of triggering this adverse event. This report calls attention to a potential problem of particular concern, that of herbal-drug interaction. This problem is not widely recognized and can cause treatment failure in patients without an immediately obvious clinical modification.This is the case of a 56 year old Caucasian man from Montenegro whose cancer history began in April 2009 after the appearance of progressive abdominal pain due to a retroperitoneum mass measuring 5 × 8 cm. The patient’s past medical record showed no relevant illness. He was an ex heavy smoker and did not drink alcohol or use illicit drugs. In May 2009 he underwent surgical removal of the abdominal mass achieving a complete margin negative (R0) resection. Histological findings of lesion specimens highlighted a low-grade de-differentiated liposarcoma. He then began follow-up. In November 2011 a computed tomography (CT) scan detected a new retroperitoneal lesion measuring 5 × 10 cm whose imaging features clearly suggested relapsed liposarcoma. He therefore underwent another radical surgery and the second histological examination supported the diagnosis of G2 de-differentiated liposarcoma with myxoid areas. Fluorescence in situ hybridization (FISH) analysis for oncogene murine double minute 2 (MDM2) status highlighted the amplification as highly specific diagnostic markers .After only three months, a new recurrence occurred and owing to the brief relapse-free time he was not considered suitable for additional surgery. Accordingly, he was treated with chemotherapy including three cycles of doxorubicin followed by three cycles of ifosfamide. In spite of chemotherapy, the lesion progressed rapidly reaching 16 × 10 cm and extending into the pre-aortic seat of the retro-peritoneum space. In April 2012 the patient was referred to our Institute and underwent second line chemotherapy with trabectedin at a dose of 1.5 mg/m [Aronia melanocarpa) daily during the last course of trabectedin and in the 2 weeks after.As the patient had this unusual muscular side effect after the fourth cycle of trabectedin, we explored possible sources that could trigger the potential trabectedin-induced rhabdomyolysis. He did not report any trauma or muscular overexertion. Due to the absence of co-morbidity, the patient had not received any other drugs during treatment with trabectedin. After further questioning in a non-threatening fashion, the patient informed us that he had been taking a commercial folk medicine preparation of chokeberry juice  extract derived from the black fruit of this shrub. This berry has a long tradition in European and North American folk medicine [In our patient this “natural foe” was chokeberry (medicine . It is amedicine . Furthermedicine ,22. Sevemedicine .From a clinical-pharmacological point of view, due to the evidence concerning decreased activities of enzymatic markers of CYP after forced feeding of chokeberry juice , the advIn conclusion, our case underlines the importance of understanding rare treatment-related toxicities such as trabectedin-induced rhabdomyolysis and the possible role of the drug-drug interactions in the pathogenesis of this rare trabectedin side effect. Furthermore, in the presence of unexpected toxicity during conventional anticancer treatment, the simultaneous use by the patient of alternative drugs should be suspected. So a thorough medication history, enquiring about nutritional supplements and over-the-counter drugs, may help raise some red flags about potential interactions with chemotherapy.Written informed consent was obtained from the patient for the publication of this report.CYP: Cytochrome P450; CT: Computed tomography; MDM2: Oncogene murine double minute 2; peg-G-CSF: Pegylated granulocytes colony stimulating factor; ECOG: Eastern Cooperative Oncology Group; CPK: Creatine phosphokinase; LDH: Lactate dehydrogenase; ALT: Alanine transaminase; AST: Aspartate transaminase.The authors declare that they have no competing interests.SS, MG and VL treated and clinically evaluated the patient, SS and MG drafted the manuscript and were principle authors of the paper, VL and AA contributed to draft the manuscript. All authors contributed to manuscript revision and approved the final manuscript.The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1472-6882/13/199/prepub


The persistence of latent HIV-infected cellular reservoirs represents the major hurdle to virus eradication with highly active anti-retroviral therapy (HAART), since latently infected cells remain a permanent source of viral reactivation. HIV establishes a persistent infection in CD4+ T lymphocytes (and to a lesser extent in macrophages as well), creating a persistent reservoir consisting mainly of latently infected resting memory CD4+ T cell. Although pre- and post-integration latencies have been described in HIV-1, the reservoir that appears to be the major barrier to eradication is composed of latently infected cells carrying an integrated provirus that is transcriptionally silent.It has been suggested that reactivation of the latent reservoirs could allow effective targeting and possible eradication of the virus. Immunoactivation therapy to reduce the latent pool of HIV by treatment with the anti-CD3 antibody OKT-3 alone or in combination with interleukin-2, substantially failed to significantly decrease the viral reservoir. Non-specific T-cell activation may induce high-level viral replication above a level that can be fully contained by ART, while increasing the susceptibility of uninfected cells.in vitro through the induction of the Janus kinase–signal transducer and activator of transcription (JAK–STAT) signalling pathway. The use of different chemical compounds targeting the PKC signalling pathway  has also been proposed as a means of reactivating viral reservoirs. Finally, HDAC blocking is an attractive potential means of inducing broad reactivation of HIV–1 reservoirs, and promising results have been achieved using the HDAC inhibitor vorinostat.Selective targeting of HIV provirus via agents that induce the expression of quiescent HIV, but have limited effects on the uninfected host cell is an alternate approach to attack latent HIV. Activation from latency to completion of the replication cycle should result in lytic cell death of CD4+ T cells. Multiple mechanisms that contribute to the maintenance of proviral latency could be targeted to activate the latent virus. As examples of potentially useful agents, IL–7 can reactivate HIV–1 in latently infected cells


This offers a good opportunity to attain biological parenthood to individuals with gonadal insufficiency including cancer survivors. However, role of various intra- and extra-ovarian factors during PF growth initiation still remain poorly understood. Ovarian biology has assumed a different dimension due to emerging data on presence of pluripotent very small embryonic-like stem cells (VSELs) and ovarian germ stem cells (OGSCs) in ovary surface epithelium (OSE) and the concept of postnatal oogenesis. The present study was undertaken to decipher effect of follicle stimulating hormone (FSH) and basic fibroblast growth factor (bFGF) on the growth initiation of PF during organ culture with a focus on ovarian stem cells.Cryopreserved ovarian cortical tissue acts as a source of primordial follicles (PF) which can either be auto-transplanted or cultured Serum-free cultures of marmoset (n=3) and human  ovarian cortical tissue pieces were established. Cortical tissue pieces stimulated with FSH (0.5 IU/ml) or bFGF (100 ng/ml) were collected on Day 3 for histological and molecular studies. Gene transcripts specific for pluripotency , early germ cells  and to reflect PF growth initiation  were studied by q-RTPCR.A prominent proliferation of OSE (which harbors stem cells) and transition of PF to primary follicles was observed after FSH and bFGF treatment. Ovarian stem cells were found to be released on the culture inserts and retained the potential to spontaneously differentiate into oocyte-like structures in extended cultures. q-RTPCR analysis revealed an increased expression of gene transcripts specific for VSELs, OGSCs and early germ cells suggestive of follicular transition.in vitro. Results provide a paradigm shift in the basic understanding of FSH action and also offer a new perspective to the field of oncofertility research.The present study shows that both FSH and bFGF stimulate stem cells present in OSE and also lead to PF growth initiation. Thus besides being a source of PF, cryopreserved ovarian cortical tissue could also be a source of stem cells which retain the ability to spontaneously differentiate into oocyte-like structures  This approach has also been implicated as a possible option to achieve biological parenthood in women cancer survivors and cryopreservation of ovarian cortical tissue is currently performed prior to cancer treatment for preservation of their fertility [in vitro maturation of PF from cryopreserved tissue is an excellent option for such females and also for those who wish to delay pregnancy for career or other reasons.Culture of ovarian cortical tissue is carried out to allow the transition of resting phase primordial follicles (PF) to primary and secondary follicular stage. This is an interesting ertility -4. Currein vitro maturation of cumulus-oocyte-complexes [Ovarian cortical tissue culture however, cannot support follicle development of all stages, as the concentration and optimal timing of nutrient/growth factor requirements is stage-specific and not yet deciphered completely. Hence, a two-step procedure for follicular maturation has been proposed. The first stage involves serum-free culture of cortical tissue for 3-6 days, wherein the PF transitions to primary and secondary stage. This would be followed by omplexes ,3. Recenomplexes .Till date, many intra- and extra-ovarian factors required for PF activation and follicle development in organ cultures have been studied. Role of follicle stimulating hormone (FSH), an extra-ovarian factor, during PF transition remains controversial. It has emerged as a survival factor  however Another important factor attributed to PF transition is basic fibroblast growth factor (bFGF). It is expressed in the ooplasm of primordial and primary follicles and also in the ovarian surface epithelium (OSE), smooth muscle cells surrounding blood vessels and corpus luteum . Basic Fin vitro effect of FSH and bFGF on PF growth with a focus on ovarian stem cells in the ovarian cortical tissues of marmoset and human ovaries. Day 3 (D3) was chosen as the time point for various analyses in the present study, since available studies suggest that spontaneous activation of PF occurs within 2-3 days in vitro[in vitro.It was interesting to observe that FSH receptors and bFGF are expressed in the ovary surface epithelium ,16 which in vitro-28. HistThe study for the use of marmoset ovaries was approved by the Institute Animal Ethics Committee of National Institute for Research in Reproductive Health (NIRRH). Use of human ovarian tissue for research was approved by the Human Ethics Committee of NIRRH and King Edward Memorial (KEM) Hospital. Ovarian tissue was collected from marmosets  and women  for the study. The young ovarian sample was collected at the time of autopsy from the Surgical Pathology unit and the peri-menopausal ovarian sample was procured from the Obstetrics and Gynecology unit, KEM Hospital. All the culture studies and further analysis were carried out at NIRRH.3 size) were placed on 0.4 μm Millicell-CM inserts  fitted within six-well plates  and cultured at the interface between air and 1.1 ml of culture media, similar to that described previously [2 at 37°C and were monitored regularly under an inverted microscope with Hoffman optics . The cortical tissue pieces were collected from the cell culture inserts after three days and processed for histology and RNA extraction. The peri-menopausal ovarian tissue was treated only with FSH due to sample limitations. Details of various studies undertaken are shown in Table Ovarian tissue was transported in 0.9% normal saline-containing antibiotics  at ambient temperature to the laboratory. It was gently rinsed several times in calcium and magnesium-free Dulbecco’s phosphate-buffered saline  containing antibiotics. Ovarian cortical tissue .Levels of transcripts for pluripotent stem cells , early germ cells/oocyte specific  and to reflect PF transition into primary follicles  were analyzed by q-RTPCR. Both total Oct-4 (comprising various isoforms) and Oct-4A (transcript specific for pluripotency) were studied to arrive at meaningful data showing the presence of VSELs and differentiated OGSCs respectively, as reported earlier .Total RNA was extracted from the ovarian tissue pieces collected on D3 using standard TRIZOL (Invitrogen) method. RNA extraction method and cDNA preparation procedures were similar to those described earlier . Real-tiAmplification conditions comprised of an initial denaturation at 95°C for 3 min, followed by 45 cycles of denaturation at 94°C for 30s, annealing for 30s at respective temperature followed by elongation at 72°C for 50 sec. The melt curve analysis was performed at the end of 45 cycles to determine the homogeneity of the amplified products. The Ct values generated by CFX manager software (Bio-Rad) were normalized with respect to housekeeping gene 18S rRNA for all samples, and the Delta-Delta Ct values for each gene transcript were computed manually by subtracting delta Ct value of treated sample with that of its respective untreated control. Data obtained by q-RTPCR analysis using three adult marmosets, one each of young and peri-menopausal human ovarian samples are represented individually. All reactions were carried out in duplicates and the experiments were repeated thrice to determine the reproducibility of the results.Cortical tissue pieces cultured on the membrane of cell culture inserts appeared dark under the inverted microscope Figure A-C, F-H.Regular monitoring of the released cells on the cell culture inserts showed that the epithelial cells attached to membrane surface and formed a monolayer. The stem cells (VSELs and OGSCs) appeared to grow in size and differentiate spontaneously into oocyte-like structures Figure I-K by thSections of the ovarian cortical tissue prior to culture demonstrated intact OSE layer and several intact PF Figure . The OSEThe untreated control ovarian tissue on D3 in culture exhibited disorganization of the cortical stroma. The OSE layer appeared intact and continuous comprising of short cuboidal cells whereas the PF showed no initiation of growth Figure B, C. TheIn case of young human ovarian tissue, the OSE was discontinuous (data not shown) and relatively few intact PF were observed in the cortex before initiation of culture Figure A. Some dLevels of transcripts for pluripotent stem cells , early germ cells/oocyte specific  and to reflect PF transition into primary follicles (oocyte-specific Gdf-9 and Lhx8 and granulosa cell specific Amh) were analyzed. The transition specific markers were not studied in peri-menopausal tissue as this sample lacked PF. There was considerable biological variation in q-RTPCR results between samples but the basic trend remained similar while comparing data obtained in marmosets with that from human samples. Results were represented as fold change values over untreated control taken as 1 after normalizing with housekeeping gene (18S rRNA gene) and graphs were plotted on a logarithmic scale.Both Oct-4A and Nanog transcripts showed increased expression in marmoset and human cortical tissues treated with FSH and bFGF , proliferation of the OSE cells along with a release of stem cells onto the cell culture inserts was observed in response to FSH and bFGF. Also an increased expression of pluripotent stem cell  and germ cell  specific markers was observed. A higher fold increase of stem cell markers was observed by q-RTPCR in the young human ovarian sample. Stem cells released on the inserts retained the ability to spontaneously differentiate into oocyte-like structures as reported earlier by our group . ResultsBoth FSH and bFGF induced increase in proliferation of OSE and also increased expression of pluripotent markers Oct-4A and Nanog in all the groups including the peri-menopausal human ovarian sample devoid of PF. Expression of pluripotent stem cell markers Oct-4A and Nanog indirectly suggested the presence of pluripotent very small embryonic-like stem cells (VSELs) in the OSE as reported earlier by our group . VSELs aA similar increase in pluripotent stem cells and PF assembly in FSH analog treated mice has been reported by us . It was dark spermatogonial stem cells [Two distinct stem cell populations exist in the OSE including VSELs and their immediate descendants OGSCs . We haveem cells ,20. Thesem cells ,35.de novo PF assembly from the stem cells in response to the treatment. More studies need to be undertaken in this direction. In the present study, an increased expression of transition associated markers for oocyte (Gdf-9 and Lhx-8) and granulosa cells (Amh) were observed after the treatment, suggestive of PF transition into primary follicles, in agreement with published literature [Available literature suggests that transition of PF into primary to secondary follicles occurs within 8-20 days of ovarian cortical tissue culture. We were indeed surprised to observe varying degree of damage to the PF in the histological sections. In view of this, survival of PF in extended cultures is rather intriguing. The poor morphology could be due to compromised culture condition or a fixation artifact in the present study. But we used culture conditions based on recent studies  and stanterature -38.Stimulation of pluripotent stem and early germ cell specific markers by bFGF was not surprising since bFGF receptors are reported to be present in the OSE . ExpressThe young human ovarian sample procured from autopsy, used in the present study, showed presence of few PF whereas the stem cells survived post death and exhibited increased stimulation by FSH and bFGF treatment. A better survival of stem cells compared to the PF could be because stem cells survive hypoxic conditions better in agreement with a recent study wherein viable stem cells were isolated seventeen days post death and they retained the ability to differentiate into skeletal muscles .To conclude, culture of ovarian cortical tissue pieces is an excellent tool to study ovarian stem cell biology along with PF transition. Both FSH and bFGF stimulate the ovarian stem cells and probably aid in the assembly of stem cells as PF during extended cultures. It will be of great interest to study whether long-term culture of peri-menopausal ovarian tissue can result in PF assembly from the stem cells in response to FSH and bFGF treatment. Results of the present study have translational potential for both oncotherapy and infertility.The authors declare that they have no competing interests.SP carried out experimental work, data analysis, interpretation and manuscript drafting. DB was responsible for conceptualizing the contents of the research article, planning experiments, providing scientific inputs, data interpretation and critical drafting of manuscript. DM performed ovariectomy of marmoset samples and reviewed the manuscript. AC and AJ were responsible for providing human ovarian tissue samples and reviewing the manuscript. All authors read and approved the final manuscript.Details of markers used for characterization of pluripotent stem cells, germ cells and primordial follicle transition. Click here for file


Over the last few decades, nanocarriers for drug delivery have emerged as powerful tools with unquestionable potential to improve the therapeutic efficacy of anticancer drugs. Many colloidal drug delivery systems are underdevelopment to ameliorate the site specificity of drug action and reduce the systemic side effects. By virtue of their small size they can be injected intravenously and disposed into the target tissues where they release the drug. Nanocarriers interact massively with the surrounding environment, namely, endothelium vessels as well as cells and blood proteins. Consequently, they are rapidly removed from the circulation mostly by the mononuclear phagocyte system. In order to endow nanosystems with long circulation properties, new technologies aimed at the surface modification of their physicochemical features have been developed. In particular, stealth nanocarriers can be obtained by polymeric coating. In this paper, the basic concept underlining the “stealth” properties of drug nanocarriers, the parameters influencing the polymer coating performance in terms of opsonins/macrophages interaction with the colloid surface, the most commonly used materials for the coating process and the outcomes of this peculiar procedure are thoroughly discussed. Cancer is a leading cause of death worldwide as accounted for 7.6 million deaths  in 2008 (source: WHO Fact sheet N°297 February 2012). About 70% of all cancer deaths occurred in low- and middle-income countries. Deaths caused by cancer are forecasted to rise to over 13.1 millions in 2030 . Nevertheless, over the past few decades, significant advances have been made in fundamental cancer biology, allowing for remarkable improvements in diagnosis and therapy for cancer. Beside the development of new drugs with potent and selective activities, nanotechnology offers novel opportunities to cancer fighting by providing adequate tools for early detection and personalized treatments. Over the last decades, a number of different long circulating vehicles have been developed for theranostic purposes. These carriers are in the nanometer range size and most of them have been intended for the delivery of anticancer drugs to tissues affected by this pathology. The aim of this paper is to examine the features of “stealth” long circulating nanocarriers and the pharmacokinetic outcomes of stealthiness, and it will showcase the most investigated approaches yielding prolonged circulation of surface-engineered nanocarriers. The selective and controlled delivery of anticancer drugs to disease tissues is a requisite to prevent systemic toxicity, enhance the pharmacological profiles, and improve the patient compliance, which in turn provide for amelioration of antitumour therapy. Due to the leaky vasculature and low lymph drainage, solid tumours present erratic fluid and molecular transport dynamics. These features can yield specific accumulation of colloidal anticancer drug delivery systems into the tumour tissue by enhanced permeation and retention (EPR) effect . HoweverThe permanence in the bloodstream of nanovehicles is strongly affected by physical interactions with specific blood circulating components, opsonins. These components prevalently include complement proteins such as C3, C4, and C5, laminin, fibronectin, C-reactive protein, type I collagen, and immunoglobulins  [Poloxamines (Tetronics) and poloxamers (Pluronics) are amphiphilic block copolymers consisting of hydrophilic blocks of ethylene oxide (EO) and hydrophobic blocks of propylene oxide (PO) monomer units. Poloxamers are a-b-a type triblock copolymers (PEO-PPO-PEO) while poloxamines are tetrablock copolymers of PEO-PPO connected through ethylenediamine bridges [(PEO-PPO)O-PEO)2] –39.These polymers can be physically adsorbed on the nanocarrier surface through the hydrophobic PPO fraction .Following intravenous injection to mice and rats, poloxamer- or poloxamine-coated sub-200 nm poly(phosphazene) , PLGA naFor a given triblock polymer, it was found that both surface polymer density and coating layer thickness are affected by the particle size: smaller particles (below 100 nm) adsorb fewer polymer molecules per unit area than larger particles. Therefore, the polymer surface density decreases as the particle size decreases. Additionally, Pluronic adsorption on larger particles is relatively weaker than on smaller particles, which can affect the rate and extent of displacement of adsorbed polymers by blood components . The surface adsorption efficiency and the stability of the polymer coating are strictly related to the polymer composition, namely, PO/EO molar ratio and PPO and PEO chain length . Pluronic F-108 NF (poloxamer 338) has a bulkier central hydrophobic block and longer side hydrophilic arms  as compared to Pluronic F-68 NF . Accordingly, Pluronic F-108 NF forms more stable coating layers than Pluronic F-68 NF. In vivo, Pluronic F-68 NF-modified nanoparticles accumulate at 74% of the dose in the liver in 1 h, while the liver accumulation of Pluronic F-108 NF-modified nanoparticles was 67% .Dextran is a polysaccharide largely used for biomedical applications including for the decoration of nanoparticulate drug delivery systems . Dextran coating was found to bestow long circulating properties on liposomes . SimilarInterestingly, 70 kDa dextran coating was also found to reduce the burst of drug release from liposomes .Dextran was used to coat superparamagnetic iron oxide nanoparticles for magnetic resonance imaging , 52. ParSialic acid derivatives received considerable interest as potential materials to confer stealth properties to nanoparticles for drug delivery applications. Sialic acid is a component of eukaryotic cell surface and plays an important role in preventing the removal of self-tissue by low level of complement activation through the alternative pathway. Desialylation of erythrocyte membranes results in reduction of factor H binding on their membrane that switches them from nonactivators to activators of the alternative complement pathway , 54. PlaSurolia and Bachhawat demonstrated that liposomes coated with sialic acid derivatives are poorly recognised by the macrophages as they mimic the mammalian cell surface .Stealth nanocarriers have been obtained using a variety of polysialic acid derivatives, including gangliosides –61, gangThe liposome coating with the monosialoganglioside GM1 , a brainFew studies postulated that the shielding of the negative charges of GM1 by the bulky, neutral hydrophilic sugar moieties is paramount to its stealth activity . NeverthInterestingly, studies performed with mice and rats showed that the gangliosides have a specie-specific activity. Indeed, the GM1 decoration was effective in mice while it did not have any beneficial effect on the circulation time of liposomes in rats .Zwitterionic phospholipid derivatives have been demonstrated to reduce the complement activation induced by liposomes .Based on this evidence, synthetic zwitterionic polymers have been used to produce stealth drug delivery systems. These materials bind water molecules more strongly than polymers forming hydrogen bridges such as PEG. Furthermore, they provide electrostatically induced hydration  that decsn-glycero-3-phosphoethanolamine (poly(carboxybetaine)-DSPE) (Liposomes coated with poly(zwitterionic) 2 and 5 kDa poly(carboxybetaine)-1,2-distearoyl-e)-DSPE)  possess Poly(carboxybetaine) is more chemically stable than PEG and has lower interactions with proteins over short and long time . This maPolyglycerols (PGs) are biocompatible and flexible hydrophilic aliphatic polyether polyols, with an antifouling effect comparable to PEG . By virtLiposomes decorated with PGs exhibit extended blood circulation time and decreased uptake by liver and spleen .Self-assembled monolayers (SAMs) of dendritic PGs were deposited on gold surface through a disulfide linker group (thioctic acid). Surface Plasmon resonance (SPR) measurements showed that PGs monolayers efficiently prevent the adsorption of proteins. It was concluded that dendritic PGs behave as antiopsonic materials because they combine the characteristic structural features of several protein-resistant materials: flexible aliphatic polyether structure, hydrophilic surface groups, and a highly branched architecture . The inhSynthetic polyacrylic and polyvinyl polymers bearing hydrophobic moieties have been prepared to coat liposomes. The hydrophobic function allows for the polymer anchoring on the particle surface.Palmitoyl- or phosphatidylethanolamine- (PE-) terminated derivatives of poly(acryl amide) (PAA), poly(vinyl pyrrolidone) (PVP), and poly(acryloyl morpholine) (PAcM) have been found to exert comparable stealth effects on liposomes in vivo. This behaviour depends on the length of the hydrophobic alkyl function, the polymer molecular weight, and its surface density , 89.Comparative studies performed with palmitoyl-or PE-functionalized 6–8 kDa PAA, PVP, and PEG showed that the PEG derivative has slightly better performance as compared to the other polymers. Macromolecules containing shorter hydrophobic moieties than palmitoyl- or phosphatidylethanolamine-, namely, dodecyl alkyl chains, or higher polymer molecular weight (12–15 kDa) showed a lower effect on circulation time of liposomes. Short hydrophobic moieties cannot efficiently anchor the polymer on the liposome surface as the energy of the polymeric chain motion is higher than the energy of the anchoring alkyl chain interaction with the liposomal phospholipid bilayer , 90. The16H33–S– as hydrophobic anchor (PVA-R) on the liposome surface was directly proportional to the polymer molecular weight and to the concentration of the polymer solution used for the coating process. Furthermore, it was found that the PVA-R density on the liposome surface increased as the molecular weight of the polymer decreased. The PVA-R on liposomes was not detached by dilution or in presence of serum while preventing the adsorption of plasma proteins. In vivo the PVA-R-coated liposomes showed prolonged permanence in the circulation, which increased as the PVA molecular weight increased. The circulation time of liposomes coated with 1.3% mol of 20 kDa PVA-R was comparable to that of liposomes coated with 8% mol of 2 kDa PEG-1,2-distearoyl-sn-glycero-3-phosphoethanolamine (PEG-DSPE). Detailed investigations showed that the increased permanence in the bloodstream was strictly related to the PVA-R stability on the liposome surface that was higher compared to PEG-DSPE  in the presence of monocarboxyl-terminated PEG [Iron oxide nanoparticles stabilized by carboxyl coordination of the surface oxide molecules were prepared by high-temperature decomposition of tris(acetylacetonate) iron(III) [Fe(acac)ated PEG .Postproduction iron oxide nanoparticle decoration was performed using silane-terminating PEG. The silane group strongly interact with the oxide on the nanoparticle surface . PEGs de−1 nanoparticles) over a wide range of protein concentration (0.05 to 10 g L−1) [Phosphonic acid-terminated poly(oligoethylene glycol acrylate) [poly(OEGA)] was grafted to iron oxide nanoparticles through the phosphonic acid end group that provide strong interaction with iron oxide nanoparticles. The poly(OEGA-) stabilized iron oxide nanoparticles showed significant stealth properties and exhibited low BSA adsorption (<30 mg g0 g L−1) .3 decomposition in high-boiling organic solvents were postproduction PEGylated by the ligand exchange method. The nanoparticles produced with oleic acid, hexane, or trioctyl phosphine oxide (TOPO) coating were combined with PEG-silanes, PEG-PEI, PEG-PAMAM, PEG-fatty acid to allow for the coating exchange in aqueous medium [Iron oxide nanoparticles synthesized by Fe(acac)s medium –168.3 thereby converting the particle surface from hydrophobic to hydrophilic according to a postproduction protocol [Dopamine has been proposed as an alternative anchoring group to silane to coat magnetic nanoparticles. Dopamine has high affinity for the iron oxide and can be conjugated to PEG through the amino group. PEG-dopamine was used to displace the oleate/oleylamine coating on the particles produced by high-temperature decomposition of Fe(acac)protocol .“Growing from” approaches based on living radical polymerization techniques such as Atom-Transfer Radical-Polymerization (ATRP) and Reversible Addition-Fragmentation chain-Transfer (RAFT) polymerization have been largely investigated to coat preformed iron oxide nanoparticles with PEG copolymers. ATRP polymerization of PEG-methacrylate (PEG-MA) was performed in aqueous solvent after a silane initiator (4-(chloromethyl) phenyl trichlorosilane) immobilization on iron oxide nanoparticle surface. After poly(PEG-MA) grafting, the uptake of the nanoparticles by macrophages was reduced from 158 to less than 2 pg per cell confirming the excellent shielding capacity of this novel material .Alternatively, the ATRP polymerization of the PEG-MA was performed according to a solvent-free protocol. The macroinitiator on the surface of the magnetic iron oxide nanoparticles was introduced by exchanging the surfactant (oleic acid) on the nanoparticle surface with 3-chloropropionic acid. The exchange made the nanoparticles soluble in PEG-MA that was then polymerized by ATRP. No difference in terms of capacity to evade macrophage uptake was detected when poly(PEG-MA-) coated iron oxide nanoparticles were prepared in water or by the solvent-free method .Hyperbranched polyglycerol (HPG) has recently emerged as a biocompatible and resistant material to protein adsorption, which was ascribed to its hyperbranched nature . HPG-gra3− in solution can in fact be reduced by the amino groups of the PEI block of poly(ethylenimine)-poly(ethylene glycol) block copolymer (PEI-b-PEG) [Gold nanoparticles have been PEGylated according to “one-pot” methods. AuClI-b-PEG) .−1). Thiol-PEG can react in solution with gold nanoparticles providing colloidally stable and biocompatible gold nanoparticles [Postproduction PEGylation strategies have relied mostly on the use of thiol (-SH) terminated PEGs because of the very high specific binding affinity of thiol groups to metal gold  conjugated on gold nanoparticle surface substantially improved the stability in biological media . Gold naThe in vivo performance of gold nanorods stabilized with thiol-PEG depends on the polymer molecular weight. Accordingly, stable nanorods for blood circulation were obtained with 5 and 10 kDa PEGs while smaller or larger PEGs were poorly flexible or bend into a mushroom-like configuration, respectively , 178.2 per chain, which is comparable to the hydrodynamic size of the mPEG-thiol molecule [The maximum achievable density of PEG chains on gold nanoparticles was 2.2 nmmolecule . At satuLayer-by-layer (LBL) coating approaches relying on electrostatic interactions between polymer chains and gold nanoparticle surface have been investigated to build up a hydrophilic polymer corona on gold nanoparticles. The colloidal core of gold nanoparticles was coated with layers of poly (PAH) and poly-(styrenesulfonate) (PSS). F-HPMA, a hydrophilic terpolymer composed by 90% mol of N-(2-hydroxypropyl) methacrylamide, was then conjugated to the amino groups of PAH to yield core/shell multifunctional nanoparticles. The terpolymer provides a highly water-solvated corona layer that minimizes the opsonisation process and bestows remarkable stealth properties on nanoparticles. The multifunctional nanoparticles did not show a significant degree of adsorption on the macrophage membrane or internalization by the cells .2 and 1,2-distearoyl-sn-glycero-3-phosphoethanolamine (DSPE) were conjugated to the backbone of polyglutamic acid (PGA) at 60% and 10% mol ratio with respect to the PGA monomers, respectively. Gold nanoparticle coating was achieved by exchanging the citrate adsorbed on gold particles, obtained by tetrachloroauric acid reduction, with the multifunctional polymer PGA-DSPE-mPEG. These functionalized colloidal systems showed high stability to aggregation over 48 hours of incubation in 50% fetal calf serum [PEG was grafted on gold nanoparticle surface according to a process named physisorption. PEG-NHlf serum .N,N-dimethylamino) ethyl methacrylate (PEG-b-PAMA) was shown to improve the long-term stability of gold nanoparticles. The tertiary amino group of PAMA can strongly adsorb to the surface of gold nanoparticles even though the mechanism of immobilization is not clear yet. The alkylation of pendant amino groups along the polymer backbone seems to favour the interaction of the nitrogen atom with gold. The colloidal system was physically stable over 4 days of storage in 95% human serum [Polyethylene glycol-block-polypropyltrimethoxysilane and hydroxymethyltriethoxysilane mixtures in the presence of sodium hydroxide and a surfactant [Silica nanoparticles possessing an organosilica core and a PEG shell were prepared according to a one-pot procedure. The process includes the co-hydrolysis and copolycondensation reactions of rfactant .Alternatively, silica nanoparticles were also PEGylated by a postproduction procedure by mesoporus silica nanoparticle reaction with PEG-silanes. It was reported that the PEG coating inhibits the nonspecific binding of human serum proteins to PEGylated silica nanoparticles. This is a guarantee if the molecular weight of the polymer is higher than 10 kDa and the polymer density (defined as wt% of the coating on the mesoporous silica nanoparticles) is 0.75 wt% and 0.075 wt% for PEG 10 kDa and PEG 20 kDa, respectively. The human serum albumin adsorption was only 2.5 wt% when PEGylated silica nanoparticles were tested compared to 18.7% for non-PEGylated nanoparticles .PEG coating on silica nanoparticles can also be achieved via electrostatic adsorption of polyethyleneimine-polyethylene glycol (PEI-PEG) copolymer. The polymeric coating was stable and tightly associated with the particle surface by virtue of the strong electrostatic interactions between the polyamino backbone of the copolymer and the negatively charged silica surface. The PEI-PEG copolymer investigated had 34 PEG chains (5 kDa) per PEI chain. The efficiency of the PEG coating in preventing the adsorption of serum proteins on the nanoparticle surface was remarkably high. Protein adsorption was at the limit of sensitivity for X-ray photoelectron spectroscopy (XPS) detection and no aggregation was observed for the coated nanoparticles . The synthesis of PEO on silica nanoparticles has also been performed resulting in a 40 wt% of grafted PEO. The method has been carried out first by a two-step conjugation process of prehydrolyzed 3-glycidoxypropyl trimethoxysilane and aluminium isopropoxide to the particle surface. The subsequent polymerization of ethylene oxide was carried out at 55°C. The density of the polymer chains was found to be strictly dependent on the conjugation efficiency of the metal alkoxide on the particle surface , 187.The therapeutic advantages of nanotechnology-based drug delivery systems include improved drug bioavailability, extended duration of action, reduced frequency of administration, and lower systemic toxicity with beneficial effects on the patient acceptance. The medical management of malignancies has already benefited from the outcomes of few nanotechnology-based delivery systems. However, following intravenous administration, drug-loaded nanocarriers are rapidly opsonised by a variety of proteins, most of them belonging to the complement system, and undergo very rapid clearance via the MPS cells. In this paper, the main aspects of polymer coating technology applied to colloidal drug delivery systems have been reviewed. A number of studies and examples reported in the literature showing that stealthiness can be conferred to nanocarriers by a proper formulation design and predicated by precise physicochemical determinants have been detailed and critically discussed. The evidence reported in the literature shows that the residence time in the blood of nanocarriers can be prolonged by surface coating with neutral or zwitterionic polymers characterized by high hydrophilicity and high flexibility. Furthermore, the stealth character of the nanocarriers depends on the polymer organization on the particle surface, namely, density, thickness, and association stability. The beneficial effect of nanocarrier polymer coating in promoting stealth properties generates predominantly from the polymer ability to confer a physical barrier to the biorecognition of adsorbed opsonins by macrophages. On the other hand, the paper underlines that the components of the hydrated polymeric corona are not completely inert to the biological environment and these materials do not totally prohibit the protein opsonisation . In conclusion, while many discoveries in the field of nanotechnology have allowed to clearly improve the performances of stealth nanocarriers, a significant amount of work needs to be done before these systems achieve the required level of safety for use in humans. Studies are required to fully profile at the molecular level the interactions of the nanocarriers with the biological environment and the MPS cell response that is triggered upon contact with a specific nanocarrier.


There is not enough evidence to support the use of combined Implantable Cardiac Defibrillators (ICD) and Cardiac Resynchronization Therapy (CRT) devices in all CRT candidates. Stronger predictors of malignant ventricular arrhythmia (VA) and sudden cardiac death (SCD) are required. The purpose of this study was to assess the value of myocardial scar extension and scar characterization (core and border zone) to predict VA in CRT patients.A standard Contrast-enhanced Cardiac Magnetic Resonance (ce-CMR) was performed pre-implantation of CRT device in 78 cardiac heart failure patients (64±11 years) with ischemic and non-ischemic dilated cardiomyopathy. Systolic function was assessed by cine imaging. Myocardial scar was characterized by a standard 2D segmented gradient echo inversion-recovery sequence applied in sequential short axis slices. Total scar was expressed as percentage of LV mass and the core and border zone as percentage of total scar). Patients were followed every 6 months to look for appropriate ICD therapy episodes.th-75th=15-34 months), appropriate therapy of ICD devices occurred in 11.5% of patients. The analysis showed that the percentage of total scar was a strong predictor of VA (ventricular fibrillation or ventricular tachycardia) needing ICD therapy . In a Cox regression model adjusted for systolic function, etiology and type of prevention, the predictive value of the size of the scar remained unchanged . The percentage of border zone of the scar was also associated with the occurrence of VA . However, the proportion of core was not significant (p = 0.662). The area under the ROC curve of total scar to predict VA was 0.94, and a cut-off value of <16% of LV mass had a 100% negative predictive value to exclude VA during follow-up.During a mean follow up of 25 months (percentile 25In TRC candidates with dilated cardiomyopathy of any etiology the extent of myocardial scar and the proportion of border zone predict the occurrence of VA. Assessment of scar burden and scar characterization using ce-CMR may be helpful to decide when combined ICD/CRT devices are indicated among CRT candidates.


Patients are susceptible for knee osteoarthritis (KOA) with increasing age and obesity and KOA is expected to become a major disabling disease in the future. An important feature of KOA on magnetic resonance imaging (MRI) is changes in the subchondral bone, bone marrow lesions (BMLs), which are related to the future degeneration of the knee joint as well as prevalent clinical symptoms. The aim of this study was to investigate the changes in BMLs after a 16-week weight-loss period in obese subjects with KOA and relate changes in BMLs to the effects of weight-loss on clinical symptoms.2, an age ≥ 50 years and primary KOA. Patients underwent a 16 weeks supervised diet program which included formula products and dietetic counselling . BMLs in tibia and femur were assessed on MRI before and after the weight-loss using the Boston-Leeds Osteoarthritis Knee Score. Response to weight-loss in BML scores was dichotomised to patients experiencing a decrease in BML scores (responders) and patients who did not (non-responders). The association of BMLs to weight-loss was assessed by logistic regressions and correlation analyses.This prospective cohort study included patients with a body mass index ≥ 30 kg/mresponders in the sum of all BML size scores whereas 130 patients (77%) deteriorated or remained stable and were categorized as non-responders. Logistic regression analyses revealed no association between weight-loss < or ≥ 10% and response in BMLs in the most affected compartment . There was no association between weight-loss and response in maximum BML score . The relationship between changes in BMLs and clinical symptoms revealed that an equal proportion of patients classified as BML responders and non-responders experienced an OMERACT-OARSI response .39 patients (23%) were classified as Weight-loss did not improve the sum of tibiofemoral BML size scores or the maximum tibiofemoral BML score, suggesting that BMLs do not respond to a rapidly decreased body weight. The missing relationship between clinical symptoms and BMLs calls for further investigation. The incidence of obesity has steadily increased over the past decades and combined with the fact that patients are susceptible for knee osteoarthritis (KOA) with increasing age and obesity KOA is expected to become a major disabling disease in the future -3. It isClinical symptoms of KOA are increased in obese individuals and it is known that weight-loss can both improve symptoms as well as decrease the risk of KOA development -7. A weiAt present, KOA is considered to be a whole organ disease that affects cartilage, bone marrow, ligaments, menisci, joint capsule etc. and this can be assessed by MRI scans using the BLOKS (Boston-Leeds Osteoarthritis Knee Score) . In geneThe aim of this study was to investigate the changes in BMLs after a 16-week weight-loss intervention in obese subjects with KOA and relate changes in BMLs to the symptomatic effect of weight-loss. We hypothesized that weight-loss would improve the overall and maximum BML scores on MRI and that the effects of weight-loss on BMLs would be related to changes in clinical symptoms.192 participants were recruited November 2007–August 2008 from the outpatient clinic at the Department of Rheumatology, Frederiksberg Hospital, Denmark, to take part in the CAROT study  . Screeni2, and show primary KOA diagnosed according to the American College of Rheumatology (ACR) criteria combining clinical symptoms as well as radiographic verification of the diagnosis  and was carried out in accordance with the Helsinki Declaration II and the European Guidelines for Good Clinical Practise.The patients followed a supervised 16-week dietary program consisting of nutritional education and a diet of normal food plus meal replacements . The details of the dietary program is described elsewhere . The dieMRI scans were obtained of the target knee (worst symptomatic knee) using a high-field MRI (1.5 T) whole body scanner  following normal procedure taking one scan at each setting. Patients were positioned supine and a send/receive flex medium or large coil was fixed to the patient’s leg. The study crew ensured that patients were scanned using the same image and coil setup over time. The following five sequences were carried out:Gradient echo-scout  12.3 ms, echo time (TE) 6.6 ms, 50° flip angle, field of view (FOV) 300 x 300 mm, matrix 256 x 256). Sagittal 3D T1-FFE gradient-echo (GRE) cartilage sequence . Sagittal multi-echo T2 weighted sequence . Sagittal multi-echo proton density (PD) weighted sequence . Coronal T1 spin echo (SE) sequence . Coronal STIR (short tau inversion recovery) sequence 3 mm slices . Scheduled time for the overall MRI examination was 37 minutes.The imaging protocol used for the BLOKS assessment in this study was the sagittal multi echo PD and T2 weighted scans, the STIR scan, the coronal SE T1 scan, as well as a 3D reconstruction of the T1-FFE scan.BMLs appear as ill-defined signal intensity changes in the subchondral bone that are hypointense on T1w images and hyperintense on STIR images . BML assInitial inter- and intra-reader analyses were performed by HG and MB by scoring 20 consecutively selected scans from the 187 completed baseline MRI examinations. Selection was performed so that the analyses were completed on 10 females and 10 males. The selected patients displayed all levels of KOA joint damage, evaluated by the medial compartment KL grade on radiographs . This procedure was similar to what was done in the original BLOKS publication; please see the online appendix . DiscrepAll imaging assessments were performed using the MacOS X based Osirix software (version 3.9.1) .Bi-plane weight-bearing semi-flexed radiographs were taken of the index knee; one in the postero-anterior and one in the lateral view . The radiographs were obtained at inclusion by the same radiographers, using a Philips Optimus apparatus and a standardized protocol through all examinations.MB analysed all radiographs using an atlas  and perfMeasurements of the tibiofemoral minimum joint space width (mJSW) in the most affected tibiofemoral compartment, judged by the KL grade, were performed by HG. Inter- and intra-reader analyses of the mJSW were performed by scoring the same scans as described above for the inter- and intra-reader analyses of BLOKS .nd metatarsal head, lateral malleolus, posterior aspect of calcaneus, lateral aspect of the leg, lateral femoral epicondyle, lateral aspect of the thigh, bilaterally on anterior and posterior superior iliac spines) according to the Plug-in-Gait biomechanical model, and anthropometric measurements  to determine joint centres. The mechanical axis alignment was defined as the frontal plane knee joint angle expressed in the local joint coordinate system. This procedure yields estimates of mechanical axis alignment similar to full-limb weight-bearing radiographs (R2=0.54) but without exposure to radiation [Mechanical axis alignment was measured using a 6 camera stereophotogrammetric system  with markers placed on anatomical landmarks  of hamstrings and the quadriceps muscles were assessed by isometric dynamometry at 60° (0° is full extension) knee joint flexion angle  as described . After cPatient reported outcomes were examined at baseline and after 16 weeks by assessing the OMERACT-OARSI Responder Criteria and KOOS pain and function in daily living (ADL), which were completed by the patients on a validated touch screen solution .The OMERACT-OARSI responder criterion was assessed by average pain, disability and patient global assessment of disease impact during the last week using 0 to 100 mm visual analogue scales (VAS) . The Kneper protocol analysis). The most affected knee compartment was the one with the worst status on radiographs and only this single compartment for every patient case was included in the analyses exploring the relationship between weight-loss and BML responses. In order to examine the importance of changes observed in BML gradings we calculated the least significant criterion (LSC) using the following formula LSC= 1.96 * SE * √ 2 * (1-r)  [Analyses were performed on available cases (elation) . To over16 - x0)/x0 * 100%). Regarding the influence of weight-loss on BMLs we conducted both regression and correlation analyses. A logistic regression model analysed the influence of a weight-loss < or ≥ 10% of the initial body weight on the response in BML scores. These regressions were repeated with adjustments for age, gender and group allocation in the underlying randomisation to either the LED or VLED intervention. The correlation analyses examined the relationship between weight-loss, as a continuous variable, and the response in BML scores. Changes in symptomatic outcomes (KOOS and OMERACT-OARI response) were analysed as relative measures, being the percentage change from baseline ((x16 - x0)/x0 * 100%), and were then analysed in relation to the dichotomised response in BML scores.We dichotomised the observed changes and our dependent variable contained 0 for patients who progressed or remained stable (non-responders) in their BML scores and 1 for patients responding positively (responders) . As a seThe inter- and intra-reader analyses of the mJSW were performed by applying an ICC two-way mixed model with measures of absolute agreement whereas the inter- and intra-reader analyses of BLOKS were performed by weighted kappa analyses based on individual scores.A P-value less than 0.05 (two-tailed) or a 95% confidence interval (CI) not including the null hypothesis was regarded as statistically significant. All data analyses were carried out applying SAS software .The study included 192 obese KOA patients and following the 16 weeks of diet intervention 175 (%) patients remained in the study. 187 (97%) MRI scans were completed at baseline, 172 (98%) MRI scans were obtained at week 16 and this left the study with 169 (97%) patients with complete MRI datasets. No statistical significant differences were detected between baseline characteristics of all the initially included patients (n=192) and the 169 patients included in the analyses in this paper (p<0.05).2 (SD 4.5) and was 62.7 years (SD 6.3) of age  and an average participant had a BMI of 37.0 kg/mge Table . One in The inter- and intra-reader analyses of the subscores in BLOKS revealed kappa values between 0.51-0.90 Table  and the responders) and 130 patients (77%) deteriorated in their score or remained stable (non-responders) (Table 39 patients (23%) experienced a decrease in the sum of all BML size scores . Adjusting for age, gender and randomization group did not change the results significantly . There were no association between weight-loss during the diet intervention period and the response in maximum BML score . Focusing the analyses to only include patients having a baseline BML score > 0 (N = 139) did not have any important impact on the results (Table There were no differences in BML responses in the underlying RCT (data not shown).responders and non-responders experienced an OMERACT-OARSI response . KOOS pain improved 23.9 and 22.6% for BML responders and non-responders, respectively  and similar results were found for KOOS ADL . Comparable results were found when analysing the association between response in maximum BMLs and KOOS pain  as well as KOOS ADL .The relationship between changes in BMLs and clinical symptoms revealed that an equal percentage of patients classified as BML In this study we aimed to assess whether or not a 16-week diet intervention program would impact on BMLs, and we assessed both the maximum and overall BML scores. Results from the study were that changes seen in the total tibiofemoral sum of BML scores and maximum BML scores were not impacted by weight loss. Also, we found no relationship between improvements in clinical symptoms and BMLs following a weight-loss.BMLs are a common finding in painful osseous conditions and have been proposed to consist of extracellular fluid that causes pain via an increased intraosseous pressure ,39. In cPrior to the intervention our cohort had a high prevalence of BMLs, which is somewhat in contrast to previous reports investigating symptomatic KOA patients ,43. In cThe present study did not provide any biological explanation for the response in BMLs after weight-loss. Data from the MOST study suggest an association between alignment of the knee and progression of BML mediated through increased mechanical load in a compartment-specific manner, since alignment is believed to influence load distribution . The preNeither did the study provide data clarifying the relationship between BML changes and the effects of weight-loss on clinical symptoms. Several factors are likely to have an impact on the assessments and interpretations of clinical symptoms in KOA. First of all, longitudinal changes in BMLs have not been shown to correlate with corresponding changes in KOA symptoms . SecondlThe inter- and intra-reader analyses of both the BLOKS assessments and the mJSW showed results that were comparable to those achieved by others ,53,54.The study has some limitations. The agreement on when an observed change is important is not yet clarified for BMLs assessed by BLOKS, but we believe that our estimate of the LSC validated our results. Our analyses are based on single determinations of MRI items which may result in erroneous estimations of associations due to the subjective variability of these. The MRI protocol for this study did not include all the recommended sequences for optimal gradings of KOA pathology. However, we believe that the BLOKS assessment performed in general served the question addressed in this study. The protocol only included coronal slices for the assessment of BMLs in trochlea and the dorsal portion of the femoral condyles and even though we consider this to be reasonable ,56 it isWeight-loss did not improve the total sum of tibiofemoral BML size scores or the maximum BML score, suggesting that BMLs do not respond to a rapidly decreased body weight. Ancillary sensitivity testing that excluded patients with no BMLs at baseline showed similar results and thus supported the initial findings.In conclusion, this study did not show any association between weight-loss and response in BML scores and failed to connect a positive BML response to clinical improvements. The missing relationship between changes in clinical symptoms and BMLs calls for further investigation.ACR: American college of rheumatology; BLOKS: Boston-Leeds Osteoarthritis Knee Score; BMI: Body mass index; BML: Bone marrow lesion; CAROT: Influence of weight-loss or exercise on cartilage in obese knee osteoarthritis patients: a randomized controlled trial; KOA: Knee osteoarthritis; FOV: Field of view; GRE: Gradient echo; ICC: Intraclass correlation coefficient; KL: Kellgren & Lawrence grade; KOOS: Knee Injury and Osteoarthritis Outcome Score; LED: Low energy diet; LSC: Least significant criterion; mJSW: Minimum joint space width; MPR: Multi planar reconstructed; MRI: Magnetic resonance imaging; MVC: Maximal voluntary contraction; OR: Odds-ratio; PD: Proton density; SE: Spin echo; STIR: Short tau inversion recovery; TE: Echo time; TR: Repetition time; VAS: Visual analogue scale; VLED: Very low energy diet.HB, RC and MH received travel grants to attend scientific meetings from the Cambridge Manufacturing Ltd. The sponsor had no involvement with respect to design, collection of data, analysis, interpretation, writing or submission.HG made all the analysis and interpretation of data, drafted the manuscript and approved the final version. MB contributed to the conception and design, analysed all MRI and radiographs, revised the manuscript several times and approved the final version. RC contributed to the conception and design, especially the statistics, revised the manuscript and approved the final version. EMB, MH and BDS contributed with the overall design idea, revised the manuscript and approved the final version. HB contributed to the conception and design, revised the manuscript and approved the final version.The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1471-2474/14/106/prepub


To characterize and eliminate a new type of image artifact in concurrent transcranial magnetic stimulation and functional MRI (TMS-fMRI) caused by small leakage currents originating from the high-voltage capacitors in the TMS stimulator system.The artifacts in echo-planar images (EPI) caused by leakage currents were characterized and quantified in numerical simulations and phantom studies with different phantom-coil geometries. A relay-diode combination was devised and inserted in the TMS circuit that shorts the leakage current. Its effectiveness for artifact reduction was assessed in a phantom scan resembling a realistic TMS-fMRI experiment.The leakage-current-induced signal changes exhibited a multipolar spatial pattern and the maxima exceeded 1% at realistic coil-cortex distances. The relay-diode combination effectively reduced the artifact to a negligible level.The leakage-current artifacts potentially obscure effects of interest or lead to false-positives. Since the artifact depends on the experimental setup and design , we recommend its assessment for each experiment. The relay-diode combination can eliminate the artifacts if necessary. J. Magn. Reson. Imaging 2009;29:1211–1217. © 2009 Wiley-Liss, Inc. COMBINING TRANSCRANIAL MAGNETIC STIMULATION (TMS) ,2 with fnot being discharged, since the electronic switch that controls the discharging has a finite resistance. This leakage current through the TMS coil generates a local magnetic field distortion, and hence EPI geometric and intensity distortion  currents in the TMS coil, present even when no TMS pulse is applied. This is due to the following effect: In general, TMS stimulators make use of high-voltage capacitors for creating strong currents through the TMS coil when they are discharged. However, the high voltage can also drive a small leakage current even when the system is stortion . Here westortion .TMS flowing through the TMS coil will generate a magnetic field TMS∝ITMS. For small TMS compared to the main field 0, the strength of the net magnetic field can be approximated as ∥Bz0 + BTMS will cause geometric distortions (mainly) in the phase-encoding (PE) direction of the EPI image and concomitant signal changes due to compression/stretching of voxels  and the PE direction either in the x (left–right) or y (anterior–posterior) direction. The long axis of the coil (connecting the two loops) pointed in the x direction. Any electric current If voxels . The magITMS were acquired for the same actual TMS coil configurations as described above for the simulations. Data were acquired with a 1.5 T whole-body MRI scanner  using the standard CP head coil and body transmit coil. TMS was conducted using a MagStim Rapid system  with a custom-built MR-compatible figure-of-eight stimulation coil : two windings of 10 turns each, inner/outer diameter winding 53 mm/86 mm, thickness of TMS coil's casing (measured as distance between outer coil surface and center of winding) ≈15 mm, coil resistance RTMS ≈100 mΩ, maximal voltage/current at 100% stimulator output ≈1.65 kV/5 kA. A dome-shaped water phantom was positioned with the phantom's flat surface parallel to the transverse plane . At ITMS = 0 mA and ITMS = 5 mA, double echo FLASH images , field of view (FOV) = 250 × 250 mmITMS as the independent variable.To improve the signal-to-noise ratio (SNR), images were spatially smoothed with an isotropic Gaussian kernel of full-width at half-maximum (FWHM) = 8 mm. To estimate the change in image intensity due to the experimentally induced current flowing in the TMS coil, SPM5 linear regression analysis  was performed on the images with In Experiment 1, controlled currents were driven through the TMS coil using an external power supply to estimate the level of current-related artifacts. In our second phantom experiment, we studied the more practically relevant case of actual leakage currents originating from the TMS stimulator itself see . Since trel) and not through the TMS coil (ITMS). While the relay is closed or its status changed, TMS pulses must not be applied, since this type of high-voltage relay does not support the high currents (in the range of kA) of a TMS pulse. Therefore, the relay and TMS stimulator were controlled via a dedicated DOMINO 1 microcontroller .The TMS setup was almost identical to that used in the previous phantom experiment. However in addition, a high-voltage relay  was introduced in parallel and a high-voltage diode  arrangement in series to the TMS coil .maximal EPI signal distortions on the order of 1% in a slice 4 cm away from the TMS coil —a realistic coil-cortex distance in TMS  direction. The geometric distortions lead to compression/stretching of the imaged voxels and subsequently to signal decreases/increases. As shown here, the TMS stimulator's high-voltage capacitors can be a source of charge-level-dependent leakage currents. Even small leakage currents of ≈1 mA as observed in our experiments can cause locally e in TMS .If the leakage currents were constant, they would only cause an offset in the signal amplitude. Such an offset would not affect the results of a conventional fMRI analysis, since it is only sensitive to signal changes over time . HoweverHere we have developed a method to minimize the leakage current flowing through the TMS coil by introducing a relay in parallel and diodes in series with the TMS coil see . When thFor brevity, we only presented data from experiments on a phantom without discharging the stimulator. However, the theory and the mechanism of the leakage currents indicate that these results readily generalize to human TMS-fMRI experiments. The effects do not depend on what object is imaged or whether TMS pulses are applied during an experiment. Further, the geometric setup used here is typical for human studies  and simulations show that other coil orientations result in artifacts of the same order of magnitude (not shown here). Most important, the relay-diode artifact suppression was successfully applied in human TMS-fMRI experiments , effectively shorting the circuit. Therefore, careful testing of possible interactions with the radiofrequency and gradient fields is advised (applying the tests described below and, eg, Ref. We recommend the assessment of this potential artifact for each individual TMS-fMRI experiment. An established test method is to run the same experiment on a phantom as used later on the human subjects (see, for example, the supplementary material in Ref. In conclusion, we have identified a new type of artifact potentially arising in concurrent TMS-fMRI experiments, caused by small leakage currents flowing through the stimulation coil. The addition of a bypass relay and diodes into the TMS circuit can efficiently eliminate any leakage current to improve image quality beneath the TMS coil, providing one useful step toward a more accurate assessment of BOLD activity close to the TMS stimulation site. Since the artifact depends on various factors, for optimal results we recommend assessment of this potential artifact for each individual TMS-fMRI experiment and application of the correction method presented here if necessary.


Alzheimer's disease is a growing concern in the modern world. As the currently available medications are not very promising, there is an increased need for the fabrication of newer drugs. Curcumin is a plant derived compound which has potential activities beneficial for the treatment of Alzheimer's disease. Anti-amyloid activity and anti-oxidant activity of curcumin is highly beneficial for the treatment of Alzheimer's disease. The insolubility of curcumin in water restricts its use to a great extend, which can be overcome by the synthesis of curcumin nanoparticles. In our work, we have successfully synthesized water-soluble PLGA coated- curcumin nanoparticles and characterized it using different techniques. As drug targeting to diseases of cerebral origin are difficult due to the stringency of blood-brain barrier, we have coupled the nanoparticle with Tet-1 peptide, which has the affinity to neurons and possess retrograde transportation properties. Our results suggest that curcumin encapsulated-PLGA nanoparticles are able to destroy amyloid aggregates, exhibit anti-oxidative property and are non-cytotoxic. The encapsulation of the curcumin in PLGA does not destroy its inherent properties and so, the PLGA-curcumin nanoparticles can be used as a drug with multiple functions in treating Alzheimer's disease proving it to be a potential therapeutic tool against this dreaded disease. Curcuma longa, a herbaceous tuberous plant endemic to South Asia. Curcumin (M.W 368.37) is chemically diferuloymethane (C21H20O6) and is a mixture of three major curcuminoids- curcumin, demethoxy curcumin and bis-demethoxy curcumin (Structure of curcumin: Alzheimer's disease (AD) is a growing concern in the modern world. It is the most common type of dementia affecting mainly the elderly population. With the better living conditions and longer life span, the number of individuals affected with AD is increasing exponentially ab initio computational study Apart from the anti-amyloid properties However, the application of curcumin is very limited due to its hydrophobic nature and non solubility in water The efficiency of the medication also depends on the bio-availability at the correct location. Targeting the central nervous system has always been a challenge due to its inaccessibility. The stringent blood-brain barrier due to the presence of tight junctions within the capillary endothelium severely restricts the delivery of therapeutics to the brain The objective of this paper is to report the conjugation of Tet-1 peptide to curcumin-PLGA nanoparticles, to study the anti-amyloid and anti-oxidative properties of the nanoparticles and to analyze the in vitro uptake of the nanoparticles.Single emulsion –solvent evaporation method, which is the best method for encapsulating hydrophobic compounds, was used for the synthesis of PLGA-curcumin nanoparticles The dry weight of the nanoparticles was measured. Yield of the nanoparticle synthesis was calculated Nanoparticles dispersed in Milli Q water was used for all the analyses. The size and morphology were characterized using electron microscopy. Transmission Electron Microscopy (TEM) images of the nanoparticles was taken in JEOL's JEM 2200 FS transmission electron microscope operating at 200 kV. The surface morphology was studied using Scanning Electron Microscopy (SEM) and Atomic Force Microscopy (AFM). The SEM (JEOL's JSM 7400 SEM) was operated at 5 kV. Tapping mode of the cantilever was used in the AFM analysis (Asylum Research's MFP-3D-CF AFM). The zeta potential of the particles was analyzed using the Zetasizer of Malvern Instruments. 3D TEM tomography was also performed to ascertain the encapsulation of curcumin inside the PLGA nanoparticles.The relative change in the weight of the nanoparticle with respect to the change in temperature was studied using Thermo-gravimetric Analysis (TGA). 15–20 mg of the dry nanoparticles was placed in a Platinum pan and heated from 30 to 800°C at a heat flow rate of 10°C/min under nitrogen spurge. Attenuated Total Reflectance - Fourier Transform Infra Red Spectroscopy (ATR-FTIR) (Perkin Elmer Spectrum 100 FT-IR system) was employed to study possible interaction studies and bonding pattern between PLGA and curcumin. The Electron Spectroscopy for Chemical Analysis (ESCA/XPS) was used to check the elemental composition and chemical states of the nanoparticles. The water-dispersed samples were dropped on clean Silicon substrate and dried under vacuum and the samples were analyzed in a Kratos-Axis X-ray photoelectron spectrometer  equipped with a monochromatic Al Kα X-ray source. Pass energy of 40 eV was used for the analyses. The elemental composition was calculated and curve-fitting performed using Spectral Data Processor v4.3 software.For enabling the neuronal targeting of the curcumin nanoparticles, we have tagged it with Tet-1 peptide.Tet-1 peptide was custom synthesized from Operon Biotechnologies. Curcumin encapsulated PLGA-nanoparticles dispersed in water was used for the attachment of Tet-1 peptide. EDC-NHS coupling was used for the attachment of the Tet-1peptide to the nanoparticles. Briefly, 100 µl of PLGA-coated curcumin nanoparticles was activated using 200 µl of 400 mM 1-Ethyl-3-[3-dimethlyaminopropyl]carbodiimide hydrochloride (EDC or EDAC) and 200 µl of 100 mM N-hydroxysulfosuccinimide (NHS). The mixture was gently shaken for one hour at room temperature. To this, 20 µl of Tet-1 peptide (1 mg/ml) was added, mixed well and kept under gentle shaking for 4 hours. After the incubation period, the nanoparticles were collected by centrifugation at 6000 rpm for 15 min and washed twice with Milli-Q water. The resultant nanoparticles were suspended in nuclease free water (1 ml) and used for further analyses.Curcumin has strong anti-oxidant activity The anti- amyloid activity of curcumin is a well established fact 5cells/well and incubated at 37°C in CO2 incubator with 5% CO2 for 24 hours before the addition of the nanoparticles. After the addition of the nanoparticles, the plates were further incubated for another 24 hours before proceeding with the particular assay. Both the assays-Alamar Blue (Invitrogen) and MTT (Sigma) were performed according to the manufacturer's protocols. In Alamar Blue assay, metabolically active cells convert the non-fluorescent resazurin to fluorescent resorufin. The amount of fluorescence produced is proportional to the percentage of viable cells. The fluorescence or absorbance can be measured at 560/590 nm (Excitation/Emission) or 570 nm respectively. In MTT assay, the respiring cells (live cells) reduce the yellow tetrazolium salt to purple formazan crystals by dehydrogenase enzymes secreted by the mitochondria of metabolically active cells. Thus the amount of formazan crystals formed is proportional to the number of viable cells. The absorbance was taken at 560 nm against a background absorbance taken at 690 nm . Untreated cells were taken as positive controls with 100% viability and cells without adding the assay reagents were used as a blank. A negative control was maintained for validation of the results and hydrogen peroxide served as the reagent for negative control The cytotoxicity profile of the nanoparticles was studied using two different assays- Alamar Blue and MTT assays. The cytotoxicity profile of PLGA nanoparticles (without curcumin), raw curcumin, PLGA-curcumin nanoparticles and PLGA-curcumin-Tet-1 nanoparticles were studied. Three different concentrations of each sample were tested in LAG cell line (mouse fibroblast like connective tissue). The cell lines were obtained from RIKEN Bioresource Center, Japan. The cells were grown in DMEM+10% FBS. Cells were seeded in the 96-well plates at a density of 5×107cells/ml. Nanoparticles (Tet-1 attached and unattached) were added to the cells and incubated for 4 hours. After the incubation period, the cells were washed twice with 1× Phosphate buffer saline (PBS) to remove all the unbound nanoparticles. The cells were trypsinized and resuspended in 500 µl 1× PBS. Cell fixation was achieved by treating the cells with 500 µl 3% paraformaldehyde for 20 minutes. After fixation, the cells were again washed with 1× PBS and permeabilized using 500 µl ice cold methanol and incubated for 10 minutes. After adding 500 µl 1× PBS, cells were again washed and finally resuspended in 1× PBS (1 ml). To reduce cell shock, all the steps were carried out in ice. Since curcumin-PLGA nanoparticles are auto-fluorescent in FITC channel, no other cell staining technique was used.Flow cytometry analysis was used to study the cellular uptake of the Tet-1 conjugated and unconjugated curcumin-PLGA nanoparticles. The experiments were conducted in GI-1 glioma cells. Cells were grown to attain a confluency of 102 incubator with 5% CO2 for uptake by the cells. The curcumin nanoparticles are fluorescent in the blue laser channel. To visualize the nucleus of the cells, the cells were stained with DAPI  nuclear stain, which could be observed under the UV laser.The uptake of the nanoparticles were studied using confocal laser scanning microscope (CLSM) coupled into an inverted microscope  and the images were recorded using a thermoelectrically cooled CCD camera (ANDOR IQ). All the images were taken using a 100× oil immersion objective lens. GI-1 cells grown in DMEM supplemented with neurotrophic growth factors to induce differentiation were seeded on glass bottom confocal plates. After 24 hours of growth, nanoparticles were added to the cells and incubated for 4 hours at 37°C in COCurcumin encapsulated PLGA nanoparticles were successfully synthesized by solvent evaporation method. Consistency was checked for different batches of the nanoparticles synthesized. (We have observed that when the concentration of curcumin was increased (more than 20 mg for 200 mg PLGA) during the nanoparticle synthesis, uncoated curcumin crystals were found among the nanoparticles on examination under SEM.) The synthesized nanoparticles were completely soluble in water and showed fluorescent properties under UV light. Raw curcumin suspended in water is not soluble and do not show any fluorescence under the UV light  and S3. Thermogravimetric analyses of PLGA nanoparticles, raw curcumin and curcumin-encapsulated PLGA nanoparticles shows that PLGA-curcumin nanoparticle follows the weight-loss pattern of PLGA nanoparticles. The weight-loss in raw curcumin occurs around 200°C, while that of the PLGA nanoparticle and PLGA-curcumin nanoparticle occurs around after 300°C. The weight loss in curcumin follows a gradual decrease while that of PLGA and PLGA-curcumin nanoparticles occur more rapidly. The raw curcumin and the curcumin loaded PLGA nanoparticles left some residual matter after the end of thermal decomposition, while the PLGA burnt completely without leaving behind any residual matters . The res2 peak in XPS (ESCA) (Tet-1 peptide was successfully attached to the PLGA-Curcumin nanoparticles by EDC-NHS coupling reaction. The attachment of Tet-1 was confirmed by NS (ESCA) . As XPS The DPPH assay was used to study the free-radical scavenging capacity of curcumin encapsulated PLGA nanoparticle in comparison with raw curcumin. The results are plotted in the graph .Raw curcumin showed more than 80% free-radical scavenging activity. Freshly prepared curcumin-PLGA nanoparticles had nearly 60% free-radical scavenging activity, and the anti-oxidant activity was decreased with the long term storage of the curcumin-PLGA nanoparticles. Attachment of Tet-1 to the curcumin-PLGA nanoparticles did not produce any change in its anti-oxidant activity. Comparable anti-oxidant activity was exhibited by Tet-1 attached and Tet-1 unattached curcumin-PLGA nanoparticles. A decrease in the anti-oxidant activity was exhibited by Tet-1 attached nanoparticles on long term storage, similar to that exhibited by the curcumin-PLGA nanoparticles (without Tet-1). PLGA nanoparticles (without curcumin) did not show any free-radical scavenging activity. With lesser reaction time (30 minutes-1 hour), there was a decrease in overall free-radical scavenging activity. This is because; the encapsulated curcumin requires time to leach out into the DPPH solution for the scavenging activity to take place. The point to be noted is that the encapsulation with PLGA does not destroy the anti-oxidant activity of curcumin.Anti-amyloid activity of curcumin is an interesting property studied by many researchers. Although the exact mechanism of curcumin activity is not known, it is an established fact that curcumin stops the aggregation of amyloid fibrils and help in the disaggregation of the existing plaques. We have obtained similar results  when theCytotoxicity profile of the nanoparticles was studied using Alamar Blue assay and MTT Assay. As cancer cells are sensitive towards curcumin, we chose to use LAG, a normal mouse fibroblast like connective tissue for the cytotoxicity analyses. Three different concentrations of each of the test samples  were used for the studies. The respective assay reagents were added after incubating the cells and the nanoparticles for 24 hours. The fluorescence and absorbance readings were taken and the result calculated. From the results obtained, it is concluded that PLGA-curcumin nanoparticles do not possess any significant cytotoxic effect on normal cells . The celFlow cytometry experiments helped in understanding the in vitro uptake of the targeted and non-targeted nanoparticles inside the cells. GI-1 glioma cells were used for the study. The uptake study was based on the fluorescence intensity of the curcumin nanoaprticles, which are fluorescent in the FITC channel. It was observed that Tet-1 targeting increased the neuronal uptake of the curcumin-PLGA nanoparticles compared to the non-targetd curcumin-PLGA nanoparticles. In vitro imaging studies is an important tool in studying the targeting properties of nanoparticles. Since our curcumin nanoparticles are auto-fluorescent, it provides a greater opportunity to study its movement in vitro. Curcumin nanoparticles are visible under the blue laser in the Confocal Laser Scanning Microscope (CLSM). We have conducted the cell imaging studies in GI-1 glioma cells as a model for neuronal cells. GI-1 cells were grown in DMEM supplemented with neurotropic growth factors to reduce division and facilitate differentiation to a small extend. After incubation with the nanoparticles, the cells were washed with PBS and counter stained with DAPI nuclear stain and viewed under the CLSM. The nanoparticles are taken up inside the cells very quickly and get distributed almost equally inside the cells . After tAlthough curcumin is a promising compound with numerous beneficial properties, its utility is greatly restricted by its insolubility in water. Water solubility is one of the important requirements for an ideal drug. We have achieved water solubility by preparing nanoparticle by encapsulating curcumin in PLGA. Apart from the complete solubility in water, these particles also exhibit fluorescent properties. The nanoparticles are also monodispersed in water which also adds to its advantage. According to Lockman et al., The anti-oxidant property of curcumin is one of the most important features which make its therapeutic value very high. In AD, formation of amyloid plaques results in severe oxidative stress in the neurons. The severe oxidative stress is also a reason for the neuronal loss which occurs during AD. The PLGA coated curcumin nanoparticles conserve the anti-oxidant property and is able to scavenge the free-radical source- DPPH on reacting with it. Although the nanoparticles are not as potent as raw curcumin, it does exhibit comparable reactivity. Attachment of Tet-1peptide to the nanoparticles did not affect its antioxidant activity.in vivo mice models, curcumin was able to bind to amyloid plaques and dramatically clear them. They have also reported that curcumin was able to reduce the appearance of new plaques in these transgenic mice. They suggest that apart from microglial activation, some other indirect contribution is also provided by curcumin in decreasing the amyloid burden. Similar results were published earlier by Yang et al., We were able to reproduce the amyloid binding activity shown by raw curcumin in our PLGA coated curcumin nanoparticles also. The curcumin nanoparticles were able to stop the aggregation of the amyloid plaques and disrupt the aggregates which were already formed. Earlier it was reported by Garcia-Alloza and coworkers The delivery of the drug to the right target is as important as its activity. Targeting nanoparticles to treat diseases of cerebral origin are very difficult due to the presence of the blood-brain barrier. Blood-brain barrier restricts the entry of almost all particles. Due to the unique properties of curcumin, it is able to cross the blood-brain barrier in vitro study suggest that Tet-1 targeted PLGA coated curcumin nanoparticles can be potential tool in treating AD with respect to its anti-amyloid property and anti-oxidant property, which we have reported here. Tagging the PLGA- curcumin nanoparticle with Tet-1 neuropeptide greatly increases its in vitro neuronal targeting efficiency. Although further conclusions can only be drawn only after a thorough in vivo study, the results of this preliminary study suggest the potential role curcumin can play in treating AD and the effect of Tet-1 peptide in neuronal targeting of the nanoparticles. In addition to the biocompatible nature of curcumin, the long history of its therapeutic use and low cost of the material also adds on to the advantages and makes it a promising candidate drug for treating AD.Treatment of Alzheimer's disease needs to take care of the multifactorial nature of the disease and the use of multi-functional drugs has been identified as the need-of the hour. Apart from the amyloid pathology, oxidative stress, inflammatory stress, tau protein hyperphosphorylation and cholesterol homeostasis plays important role in the development of the disease. Curcumin with its multiple properties can be a powerful drug in treating AD. The results of our Figure S1Structure of curcumin (A) and PLGA (B).(TIF)Click here for additional data file.Figure S2Solubility of curcumin under visible light: A- Curcumin dissolved in water, B- Curcumin-PLGA nanoparticles in water, C- Raw curcumin dissolved in acetone.(TIF)Click here for additional data file.Figure S3Solubility of curcumin under UV: D- Curcumin dissolved in water, E- Curcumin-PLGA nanoparticles in water, F- Raw curcumin dissolved in acetone.(TIF)Click here for additional data file.Figure S4XPS spectra of Si substrate used for the analysis of the samples. The peak at 531 represent Oxygen (1 s) peak, 285 represent Carbon peak (1 s) and the peaks at 151 and 99 represent the 2 s and 2p peaks of Silicon respectively.(TIF)Click here for additional data file.